EP3576625A1

EP3576625A1 - Sample collection tube with preservative

Info

Publication number: EP3576625A1
Application number: EP18747249.3A
Authority: EP
Inventors: Brad HUNSLEY; John Noble
Original assignee: Streck Inc
Current assignee: Streck Inc
Priority date: 2017-02-03
Filing date: 2018-02-05
Publication date: 2019-12-11
Also published as: CN110520045A; EP3576625A4; WO2018145005A1

Abstract

A blood collection sample tube including a thermoplastic polymeric material having a moisture barrier and low moisture absorption rate, optical transparency to enable viewing a sample within the tube and chemical resistance; and optionally a transparent silicon-containing coating on a majority of the side wall of the tube; and an elastomeric stopper. Prior to collecting a blood sample, the hollow chamber of the tube is in an evacuated condition relative to an ambient pressure, and the hollow chamber is partially filled with a reagent in an initial state, and the reagent is capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C.

Description

SAMPLE COLLECTION TUBE WITH PRESERVATIVE

FIELD

[0001] The present teachings relate generally to containers, and more particularly to polymeric tubes for biomedical sample collection, storage, transport and analysis.

BACKGROUND

[0002] There is a need for biomedical sample collection tubes which meet certain performance standards and are suitable for use in biomedical testing and analysis. There is a need for biomedical sample collection tubes which do not interact or alter any reagent enclosed in the tube. There is a further need for biomedical sample collection tubes which do not alter certain critical properties of the intended biomedical sample. There is a need for biomedical sample collection tubes which maintain a vacuum for a prolonged period of time, especially in conditions that the sample collection tube may experience during shipping. Such needs are particularly acute in blood sample collection and analysis.

[0003] One approach to sample tubes for blood collection has been to employ glass tubes. While useful, the risk of breakage during collection, storage and transport makes the use of glass tubes an undesirable option. An additional risk is glass delamination, in which glass particles detach from the inner surface of the tube. Another approach has been to employ polymeric collection tubes. However, the polymeric material may interact with and/or alter the enclosed reagents and/or collected blood sample. Polymeric containers may be coated with a coating covering the interior surface of the container. For example, the coating may be deposited onto the polymeric substrate by plasma enhanced chemical vapor deposition (PECVD). PECVD is a process used to deposit films from a gas state to a solid state on a substrate. Examples of polymeric containers with coatings include those in U.S. Pat. Nos. 7,985, 188, 8,834,954, 8,512,796, 9,878, 101 and those in U.S. Published Applications 20130209766, 20140295053, 20150293031 , 20140251859, 20160015600, 20140274825, 20160017490, 20150297800 and 20150290080 and incorporated by reference herein.

[0004] However, there is a need for an improved polymeric blood collection tube that provides for effective blood sample collection, storage, transport and analysis. There is a need for a blood collection tube that does not interact with and/or alter the enclosed

l reagents and/or collected blood sample. The present teachings meet one or more of the aforementioned needs.

SUMMARY

[0005] The present teachings meet one or more of the above needs by providing an improved polymeric tube for blood sample collection, storage, transport and analysis. The blood collection sample tube includes a tube having an enclosed base, a coextensive elongated side wall extending from the base and terminating at an open end, and defining a hollow chamber having an inner wall, the hollow chamber being configured for collecting a blood sample, at least the elongated side wall of the tube being made of a material including a thermoplastic polymeric material having a high moisture barrier and low moisture absorption rate, optical transparency to enable viewing a sample within the tube and chemical resistance; and optionally a transparent silicon-containing coating on at least a majority of the side wall of the tube; and an elastomeric stopper. Prior to collecting the blood sample, the hollow chamber of the tube is in an evacuated condition relative to an ambient pressure, and the hollow chamber is at least partially filled with a reagent in an initial state selected from a solid, a liquid, or a gel, the reagent including an optional anticoagulant, and a preservative composition adapted for stabilizing blood cells of the blood sample for enabling isolation of a nucleic acid or a rare cell/material circulating in the blood sample. The reagent is capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C.

[0006] The polymeric material may include a cyclic polyolefin. The polymeric material may include cyclic polyolefin copolymer (COC). The polymeric material may include cyclic polyolefin polymers (COP).

[0007] The tube may be part of a kit adapted for a non-invasive testing of a maternal blood sample of a pregnant woman. The elongated side wall of the tube may terminate at the open end in an annular rim that is optionally enlarged relative to the sidewall and is otherwise configured so that in combination with the stopper, the stopper is restricted from separation from the tube when subjected to conditions simulating a 95 kPa pressure differential for at least 30 minutes, a temperature below about 30 °C, or both; wherein the evacuated condition of the tube is capable of being maintained for at least 24 months from date of an initial evacuation of the tube. It is also important that the structure of the tube, including its closure mechanism promotes maintaining the necessary pressure within the tube, despite fluctuations in the environment during tube transport. Such fluctuations may include changes in temperature, changes in air pressure, changes in humidity and other environmental factors. More specifically, the tube must remain securely closed despite decreased humidity and decreased air pressure that may occur at higher altitudes.

[0008] The stopper may include bromobutyl rubber. The stopper of the tube may include a silicone oil coating over at least a portion of its outer surface that contacts the inner wall of the tube. The base may include a recessed dimple.

[0009] At least the inner wall of the tube may taper in diameter towards a base. The tube may have an outer diameter, as measured at the coextensive elongated side wall adjacent the open end, to length (D x L) dimension of about 13 mm x 75 mm. The tube may have an outer diameter, as measured at the coextensive elongated side wall adjacent the open end, to length (D x L) dimension of about 16 mm X 100 mm.

[0010] The tube may include a reagent fill tolerance volume of about 54 μΙ to about 66 μΙ. The tube may include a reagent fill tolerance volume of about 60 μΙ. The tube may include a reagent fill tolerance volume of about 162 μΙ to about 198 μΙ. The tube may include a reagent fill tolerance volume of about 180 μΙ. The tube may include a reagent fill by weight of plus or minus 10% of 0.0708g. The tube may include a reagent fill by weight of plus or minus 10% of 0.224g. The tube may include a draw tolerance of about 3 ml to about 5 ml. The tube may include a draw tolerance of about 4 ml. The tube may include a draw tolerance of about 7 ml to about 13 ml. The tube may include a draw tolerance of about 9 ml.

[0011] The preservative composition may contain less than about 20 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition. The preservative composition may contain less than about 15 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition. The preservative composition may contain less than about 10 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition. The preservative composition may contain less than about 5 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition. The preservative composition may contain at least about 0.1 parts per million (ppm) to about 20 ppm of an aldehyde per 100 μΙ of the preservative composition. The preservative composition may contain at least about 0.5 parts per million (ppm) to about 15 ppm of an aldehyde per 100 μΙ of the preservative composition. The preservative composition may contain at least about 1 parts per million (ppm) to about 10 ppm of an aldehyde per 100 μΙ of the preservative composition. The aldehyde may include formaldehyde.

[0012] The polymeric material may have a moisture vapor transmission rate of about 0.023 g ^■ mm/ m^{2 ■} d to about 0.045 g ^■ mm/ m^{2 ■} d at 23 °C and 85% relative humidity, as measured by DIN 53 122. The polymeric material may have a tensile strength of about 60 MPa to about 63 MPa, as measured by ISO 527, parts 1 and 2. The polymeric material may have a tensile modulus of about 2300 MPa to about 2600 MPa, as measured by ISO 527, parts 1 and 2. The polymeric material may have an impact strength (Charpy Impact Unnotched) of about 20 kJ/m2 as measured by ISO 179/1 ell. The polymeric material may have a light transmission of at least about 90%. The polymeric material may have a mold shrinkage of about 0.1 % to about 0.7%. The tube may have an average wall thickness along its length of about 2.0 mm. The coating may have an average thickness throughout the tube of about 100 nm to about 500 nm. The polymeric material may have a birefringence of less than 20 nm according to ASTM D4093-95. The polymeric material may have a glass transition temperature of about 78 °C to about 136 °C as measured by differential scanning calorimetry (DSC).

[0013] The polymeric material and coating may each have coefficients of thermal expansion that are within about twenty percent of each other. The polymeric material may include a homopolymer or copolymer that includes polyethylene, polypropylene or both. The polymeric material may include a cyclic moiety. The polymeric material may include a polyester.

[0014] The preservative may be selected from the group consisting of formaldehyde, diazolidinyl urea (DU), imidazolidinyl urea (IDU), dimethylol urea, 2- bromo-2-nitropropane-1 ,3-diol, 5-hydroxymethoxymethyl-1 -aza-3,7-dioxabicyclo

(3.3.0)octane and 5-hydroxymethyl-1 -aza-3,7-dioxabicyclo (3.3.0)octane and 5- hydroxypoly [methyleneoxy]methyl-1 -aza-3,7-dioxabicyclo (3.3.0)octane, bicyclic oxazolidines, DMDM hydantoin, sodium hydroxymethylglycinate, hexamethylenetetramine chloroallyl chloride, biocides, a water-soluble zinc salt and any combination thereof. The anticoagulant or enzyme inhibitor may be EDTA. The tube may be suitable for storing a blood sample for a period of at least about 14 days.

[0015] A blood sample stored in the tube having a silicon containing coating may result in a substantial increase in the amount of recoverable nucleic acid or rare cells (e.g., DNA, RNA, extracellular vesicles, circulating tumor cells, circulating rare cells or proteins) as compared with a polymeric tube of the same polymer but with no coating and the substantial increase may be at least about 10%. A blood sample stored in the tube having a silicon-containing coating may result in a substantial increase in the amount of recoverable nucleic acid or rare cells as compared with a polymeric tube of the same polymer but with no coating and the substantial increase may be at least about 20%. A blood sample stored in the tube having a silicon-containing coating may result in a substantial increase in the amount of recoverable nucleic acid or rare cells as compared with a polymeric tube with no coating and the substantial increase may be at least about 30%.

[0016] The tube may include an adhesive-backed label adhered to an outer wall of the tube, and the adhesive may be an acrylic based adhesive.

[0017] The reagent in the tube may include about 15% to about 70% by volume of the preservative composition prior to blood draw. The reagent in the tube may include about 15% to about 70% by volume of imidazolidinyl urea (IDU) prior to blood draw. The reagent in the tube include may include about 5% to about 20% by volume of anticoagulant prior to blood draw. The reagent in the tube may include about 5% to about 20% of anticoagulant prior to blood draw. The reagent in the tube may include about 1 % to about 10% of an agent in addition to the preservative and an anticoagulant prior to blood draw.

[0018] The tube may include an amount of aldehyde reaction agent for reacting with free formaldehyde present in the reagent sufficient to result in an aldehyde amount below 20 ppm per 100 μΙ of reagent prior to blood draw. The tube may include an amount of aldehyde reaction agent containing a primary amine for reacting with free formaldehyde present in the reagent to result in an aldehyde amount below 20 ppm per 100 μΙ of reagent prior to blood draw. The tube may include an amount of aldehyde reaction agent, which aldehyde reaction agent is selected from one or any combination of tris, glycine, or a derivative (e.g., a salt and/or an ester) of either or both, for reacting with free formaldehyde present in the reagent to result in an aldehyde amount below 20 ppm per 100 μΙ of reagent prior to blood draw.

[0019] The aldehyde reaction agent may be present in a ratio (by weight) relative to the preservative composition of about 1 :20 to about 1 : 1 . The aldehyde reaction agent may be present in a ratio (by weight) relative to the anticoagulant of about 1 :25 to about 5: 1 . The preservative composition may be present in a ratio (by weight) relative to the anticoagulant of about 1 : 10 to about 15:1 .

[0020] The reagent may be present in the tube as a liquid in an amount less than about 10% by volume of the tube but greater than about 0.1 % by volume. The reagent may be present in the tube as a liquid in an amount less than about 5% by volume of the tube but greater than about 0.1 % by volume of the tube. The reagent may be present in the tube in an amount less than about 3% by volume of the tube but greater than about 0.1 % by volume of the tube. The tube may include a reagent with one or more ingredients and a ratio of the one or more ingredients may be about 5: 1 to about 15: 1. The tube may include imidazolidinyl urea (IDU) and glycine and a ratio of imidazolidinyl urea (IDU) to glycine may be about 10: 1 .

[0021] The reagent may be capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C when the tube is subjected to relative humidity of up to about least about 50%. The reagent may be capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C when the tube is subjected to relative humidity of up to about 75%.

[0022] The reagent in the tube may include about 60% to about 90% by volume diazolidinyl urea (DU) prior to blood draw. The reagent in the tube includes about 15% to about 35% of anticoagulant prior to blood draw.

[0023] The teachings herein further provide for a blood collection sample tube comprising a tube having an enclosed base, a coextensive elongated side wall extending from the base and terminating at an open end, and defining a hollow chamber having an inner wall, the hollow chamber being configured for collecting a blood sample, at least the elongated side wall of the tube being made of a material including a thermoplastic polymeric material having a moisture barrier and low moisture absorption rate, optical transparency to enable viewing a sample within the tube and chemical resistance, an elastomeric stopper, and blood plasma including cell-free nucleic acids and a reagent located within the tube, the reagent including an optional anticoagulant, formaldehyde, and a preservative composition.

BRIEF DESCRIPTION OF THE DRAWINGS

[0024] FIG.1 is a front view of an illustrative example of a blood collection tube assembly of the present teachings.

[0025] FIG. 2(a) is a front view of an illustrative example of a 4 ml blood collection tube of the present teachings.

[0026] FIG. 2(b) is a close-up view of portion B of the tube of FIG. 2(a) depicting the upper portion of the tube terminating at an open end and including an annular rim.

[0027] FIG. 2(c) is a close-up view of portion C of the tube of FIG. 2(a) depicting the lower portion of the tube including a base and recessed dimple.

[0028] FIG. 3(a) is a front view of an illustrative example of a 10 ml blood collection tube of the present teachings.

[0029] FIG. 3(b) is a close-up view of portion B of the tube of FIG. 3(a) depicting the upper portion of the tube terminating at an open end and including an annular rim.

[0030] FIG. 3(c) is a close-up view of portion C of the tube of FIG. 3(a) depicting the lower portion of the tube including a base and a recessed dimple.

[0031] FIG. 4(a) is a graph depicting the draw volume results of twenty 9 ml blood collection tubes of the present teachings. FIG. 4(b) is a graph depicting the draw volume results of twenty 4 ml blood collection tubes of the present teachings.

[0032] FIG. 5 is a graph depicting cell-free DNA genome equivalents per 1 ml_ of plasma for a storage temperature study at 6 ± 2 °C for glass blood collection tubes and the blood collection tubes of the present teachings.

[0033] FIG. 6 is a graph depicting cell-free DNA genome equivalents per 1 ml_ of plasma for a storage temperature study at 18-25 °C for glass blood collection tubes and the blood collection tubes of the present teachings. [0034] FIG. 7 is a graph depicting cell-free DNA genome equivalents per 1 ml_ of plasma for a storage temperature study at 37 ± 2 °C for glass blood collection tubes and the blood collection tubes of the present teachings.

[0035] FIG. 8 is a graph depicting cell-free DNA genome equivalents per 1 ml_ of plasma for a shipping and handling study for glass blood collection tubes and the blood collection tubes of the present teachings.

[0036] FIG. 9(a) is a graph depicting oxygen transmission rate constants for polyethylene terephthalate (PET) tubes and 9 ml blood collection tubes of the present teachings. FIG. 9(b) is a graph depicting oxygen transmission rate constants for polyethylene terephthalate (PET) tubes and 4 ml blood collection tubes of the present teachings.

[0037] FIG. 10(a) is a graph depicting theoretical draw volumes for polyethylene terephthalate (PET) tubes and 9 ml blood collection tubes of the present teachings over 24 months. FIG. 10(b) is a graph depicting theoretical draw volumes for polyethylene terephthalate (PET) tubes and 4 ml blood collection tubes of the present teachings over

24 months.

[0038] FIG. 1 1 is a graph depicting moisture loss for polyethylene terephthalate (PET) tubes and the blood collection tubes of the present teachings over 28 days at 18-

25 °C, 30 ± 2 °C, and 50 ± 2 °C.

[0039] FIG. 12 is a graph depicting the results of a comparative analysis moisture vapor barrier study at 85 days at 22 °C, 30 °C and 50 °C of commercial plastic blood collection tubes and blood collection tubes of the present teachings.

DETAILED DESCRIPTION

[0040] The explanations and illustrations presented herein are intended to acquaint others skilled in the art with the teachings, its principles, and its practical application. Those skilled in the art may adapt and apply the teachings in its numerous forms, as may be best suited to the requirements of a particular use. Accordingly, the specific embodiments of the present teachings as set forth are not intended as being exhaustive or limiting of the teachings. The scope of the teachings should, therefore, be determined not with reference to the above description, but should instead be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled. The disclosures of all articles and references, including patent applications and publications, are incorporated by reference for all purposes. Other combinations are also possible as will be gleaned from the following claims, which are also hereby incorporated by reference into this written description.

[0041] This application claims the benefit of the filing date of U.S. Provisional Application Nos. 62/454,451 , filed February 3, 2017 and 62/454,460, filed February 3, 2017, the contents of these applications being hereby incorporated by reference for all purposes.

[0042] The present teachings pertain generally to an improved polymeric tube for biomedical sample collection, storage, transport and analysis. The polymeric tube is particularly attractive for blood sample collection. The sample collection tube of the present teachings may resist adherence of the contents of the collected biomedical sample to the walls of the sample collection tube. For example, the interior surface of the blood collection tube may resist adherence to blood cells of the drawn blood sample. The blood collection tube may resistance adherence of nucleic acids or other biological material (e.g. DNA, RNA, extracellular vesicles, circulating tumor cells, circulating rare cells or proteins) of the drawn blood sample to the walls of the tube.

[0043] The blood collection sample tube includes a tube having an enclosed base, a coextensive elongated side wall extending from the base and terminating at an open end, and defining a hollow chamber having an inner wall, the hollow chamber being configured for collecting a blood sample, at least the elongated side wall of the tube being made of a material including a thermoplastic polymeric material having a moisture barrier and low moisture absorption rate, optical transparency to enable viewing a sample within the tube and chemical resistance; and optionally a transparent silicon-containing coating on at a majority of the side wall of the tube; and an elastomeric stopper. Prior to collecting the blood sample, the hollow chamber of the tube is in an evacuated condition relative to an ambient pressure, and the hollow chamber is partially filled with a reagent in an initial state selected from a solid, a liquid or a gel. The reagent may include an optional anticoagulant, and a preservative composition adapted for stabilizing blood cells of the blood sample for enabling isolation of a nucleic acid or a rare cell circulating in the blood sample. The reagent is capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C.

[0044] The blood sample collection tube is a single use tube. The blood sample collection tube is non-pyrogenic and/or endotoxin free. The blood collection tube may be used to collect blood samples for analytical purposes.

[0045] The blood sample collection tube may be an evacuated direct draw blood collection tube which is used for stabilization and preservation of a blood sample and/or blood sample components. The blood sample collection tubes may be used for the stabilization and preservation of nucleic acids. The nucleic acid may include but is not limited to the following: genomic DNA, cell-free DNA, methlylated DNA, cellular RNA, mRNA and cell-free RNA. For example, the blood collection tubes of present teachings may be used for non-invasive prenatal testing (NIPT) which analyzes cell-free fetal DNA circulating in maternal blood. The prenatal tests may include fetal sex determination, genetic screening for one or more chromosomal conditions (e.g. trisomy) and prenatal DNA paternity testing. The tube may be part of a kit adapted for a non-invasive testing of a maternal blood sample of a pregnant woman.

[0046] The blood sample collection tubes may be used for the stabilization and preservation of one or more of the following: proteins, enzymes and antibodies, including with and without post-translational modifications. For example, for analysis by one or more of the following: surface flow cytometry, intracellular cytometry, ELISA-based assays and mass spectrometry. The blood sample collection tubes may be used for the stabilization and preservation of exosomes and/or microparticles. The blood sample collection tubes may be used for the stabilization and preservation of viruses. This may include viral inactivation and viral load quantification. The virus may include any of the following: DNA/RNA, stranded/double stranded and enveloped/non-enveloped. The blood sample collection tubes may be used for the stabilization and preservation of microorganisms such as bacteria, molds and yeasts. The blood sample collection tubes may be used for the stabilization and preservation of extracellular parasites and/or intracellular parasites. The blood sample collection tubes may be used for the stabilization and preservation of circulating tumor cells and tumor DNA. [0047] Evacuated blood collection tubes typically have an expiration date. An expired tube may have a decreased vacuum, resulting in a short blood draw and leading to an improper blood to reagent ratio. Thus, it is important that a blood collection tube be able to maintain a vacuum for an extended period of time. The blood collection tube of the present teachings may provide an evacuated blood collection tube which maintains a vacuum for an extended period of time (e.g. at least 24 months). Furthermore, the draw tolerance of the blood collection tube should be accurate, (e.g. +/- 10% of ml of the stated collection volume).

[0048] The blood collection tube may be adapted for used with a stopper. The stopper may include an elastomeric material. The stopper may include a butyl rubber derivative. The stopper may include a halogenated butyl. The stopper may include bromobutyl rubber. The stopper may include pharmaceutical bromobutyl rubber. The stopper may be utilized with stopper lubricant. The stopper may be coated. The stopper may be partially coated. For example, the coating may include silicone. The stopper of the tube may include a silicone oil coating over at least a portion of its outer surface that contacts the inner wall of the tube.

[0049] The present teachings contemplate a blood collection tube assembly including the blood collection tube of the present teachings and a stopper. It is contemplated that the seal or interface of the outer wall of the cap and the inner wall of the tube is such that the moisture transmission rate is substantially reduced. It is contemplated that the seal or interface of the outer wall of the stopper and the inner wall of the tube is such that the oxygen transmission rate is substantially reduced. An effective seal should provide both a moisture barrier and a gas barrier. The present teachings provide a blood collection tube assembly in which the stopper resists pull out from the opening of the tube. The present teachings provide a blood collection assembly which provide both an effective moisture transmission barrier thereby preventing moisture from escaping from inside of a tube filled with reagent and an effective oxygen transmission barrier thereby preventing penetration of oxygen into the tube.

[0050] The elongated side wall of the tube may terminate at the open end in an annular rim that is optionally enlarged relative to the sidewall and is otherwise configured so that in combination with the stopper, the stopper is restricted from separation from the tube when subjected to conditions simulating a 95 kPa pressure differential for at least 30 minutes, a temperature below about 30 °C, or both; wherein the evacuated condition of the tube is capable of being maintained for at least 24 months from date of an initial evacuation of the tube.

[0051] The reagent may be capable of retaining its initial state (e.g., liquid, gel, solid) for a period of at least one month over a temperature range of about 2 °C to about 30 °C when the tube is subjected to relative humidity of up to about least about 50%. The reagent may be capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C when the tube is subjected to relative humidity of up to about 75%.

[0052] The tube includes an enclosed base, a coextensive elongated side wall extending from the base and terminating at an open end, and defining a hollow chamber having an inner wall, the hollow chamber being configured for collecting a blood sample. The elongated side wall of the tube terminates at the open end in an annular rim. The base of the blood collection tube may be cup shaped. The base of the blood collection tube may include an indentation. The base of the blood collection tube may include a recess which projects inwardly into the tube. The upper portion of the blood collection tube may include one or more indentations along the exterior surface of the tube. The upper portion of the blood collection tube may include one or more indentations along the interior surface of the tube.

[0053] The blood collection tube may slightly taper along the length of the tube. The diameter of the tube as measured at the coextensive elongated side wall adjacent the open end may differ from the diameter of the tube as measured at the coextensive side wall adjacent the base. The diameter may differ from about .25 mm to about 3 mm. The diameter as measured at the coextensive side wall adjacent the open end may be greater than the diameter of the tube as measured at the coextensive side wall adjacent the base. The diameter as measured at the coextensive side wall adjacent the open end may be less than the diameter of the tube as measured at the coextensive side wall adjacent the base. At least the inner wall of the tube may taper in diameter towards a base. [0054] The sample collection tube's dimensions may be about 13 mm X 75 mm. The tube may have an outer diameter, as measured at the coextensive elongated side wall adjacent the open end, to length (D x L) dimension of about 13 mm x 75 mm. The sample collection tube's dimensions may be about 16 mm X 100 mm. The tube may have an outer diameter, as measured at the coextensive elongated side wall adjacent the open end, to length (D x L) dimension of about 16 mm X 100 mm. The sample collection tube's dimensions may be about 13 mm X 100 mm. The tube may have an outer diameter, as measured at the coextensive elongated side wall adjacent the closed end, to length (D x L) dimension of about 13 mm X 100 mm.

[0055] The blood collection tube may have a wall thickness of about 2 mm. The blood collection tube may have a wall thickness of about 1 .5 mm to about 2.5 mm. The blood collection tube may have a wall thickness of about 2 mm to about 3 mm. The blood collection tube may include an annular rim. The annular rim may extend about .50 mm from the exterior wall of the tube. The annular rim may extend about .25 to about .75 mm from the exterior wall of the tube. The wall thickness may vary along the annular rim . For example, the wall thickness along the annular rim of the tube may be greater (e.g. about .25 mm to about .75 mm greater) than the wall thickness along the wall of the tube.

[0056] The blood sample collection tubes may be suitable for about 4 ml to about 10 ml of blood collection. For example, a small blood collection tube may be suitable for about 4 ml of blood sample collection. A 4 ml blood collection tube may have a length as measured from the exterior of the base of the tube to the exterior of the stopper inserted at the open end of the tube of about 80 mm. The blood collection tube may have a diameter as measured across the exterior wall of the inserted stopper of about 16 mm. The outer diameter of the blood collection tube as measured at the coextensive elongated side wall adjacent base may be about 1 1 mm. The outer diameter of the blood collection tube as measured at the coextensive elongated side wall adjacent the open end may be about 12 mm. The inner diameter of the blood collection tube as measured at the coextensive elongated side wall adjacent the open end may be about 10 mm.

[0057] For example, a large blood collection tube may be suitable for about 9 ml of blood sample collection. A 9 ml blood collection tube may have a length as measured from the exterior of the base of the tube to the exterior of the stopper inserted at the open end of the tube of about 106 mm. The blood collection tube may have a diameter as measured across the exterior wall of the inserted stopper of about 17 mm. The outer diameter of the blood collection tube as measured at the coextensive elongated side wall adjacent the base may be about 13 mm. The outer diameter of the blood collection tube as measured at the coextensive elongated side wall adjacent the open end may be about 15 mm. The inner diameter of the tube as measured at the coextensive elongated side wall adjacent the open end may be about 13 mm.

[0058] The blood collection tube may include a reagent fill tolerance volume of about 54 μΙ to about 66 μΙ. The blood collection tube may include a reagent fill tolerance volume of about 60 μΙ. The blood collection tube may include a reagent fill tolerance volume of about 162 μΙ to about 198 μΙ. The blood collection tube may include a reagent fill tolerance volume of about 180 μΙ. The blood collection tube may include a reagent fill by weight of plus or minus 10% of 0.0708g. The blood collection tube may include a reagent fill by weight of plus or minus 10% of 0.224g. The blood collection tube may include a draw tolerance of about 3 ml to about 5 ml. The blood collection tube may include a draw tolerance of about 4 ml. The blood collection tube may include a draw tolerance of about 7 ml to about 13 ml. The blood collection tube may include a draw tolerance of about 9 ml.

[0059] The blood collection tube may include a reagent fill volume of about 1 ml. The blood collection tube may include a reagent fill volume of about 2 ml. The blood collection tube may include a reagent fill volume of about 1 ml to about 2 ml. For example, a 10 ml blood collection tube may include a reagent fill volume of about 1 ml to about 2 ml. For example, a 4 ml blood collection tube may include a reagent fill volume of about 1 ml to about 2 ml.

[0060] In one example, the sample collection tube is a 4 ml blood collection tube. The length of the tube may be about 75 mm. The tube may have an average wall thickness along its length of about 1 .7 mm to about 2.2 mm. The tube may have an average wall thickness along its length of about 2.0 mm. The inner diameter of the tube as measured at the coextensive elongated side wall adjacent the open end may be about 10 mm. The outer diameter of the tube as measured at the coextensive elongated side wall adjacent the open end may be about 12 mm. The elongated side wall of the tube terminates at the open end in an annular rim that may be enlarged relative to the sidewalk The inner wall of the annular rim may be curved. The outer diameter of the tube as measured at the annular rim may be about 12 mm to about 14 mm. The height of the annular rim may be about 1 .0 mm. The tube may taper in diameter to a base. The diameter of the base may be about 3 mm to about 4 mm. The diameter of the base may be about 3.5 mm. The base may include a recessed dimple. The recess depth may be about .25 mm to about .75 mm. The recess depth may be about .50 mm.

[0061] In one example, the sample collection tube is a 10 ml blood collection tube. The length of the tube may be about 100 mm. The tube may have an average wall thickness along its length of about 1 .7 mm to about 2.2 mm. The tube may have an average wall thickness along its length of about 2.0 mm. The inner diameter of the tube as measured at the coextensive elongated side wall adjacent the open end may be about 13 mm. The outer diameter of the tube as measured at the coextensive elongated side wall adjacent the open end may be about 15 mm. The elongated side wall of the tube terminates at the open end in an annular rim that may be enlarged relative to the sidewalk The inner wall of the annular rim may be curved. The outer diameter of the tube as measured at the annular rim may be about 15 mm to about 17 mm. The height of the annular rim may be about 1 mm. The tube may taper in diameter to a base. The diameter of the base may be about 3 mm to about 4 mm. The diameter of the base may be about 3.5 mm. The base may include a recessed dimple. The recess depth may be about .25 mm to about .75 mm. The recess depth may be about .50 mm.

[0062] The polymeric material may be characterized as having one or more of the following: a moisture barrier and low moisture absorption rate, purity, transparency, chemical resistance, heat resistance and strength. The polymeric material may have one or more of the following attributes: low density, high transparency, low birefringence, extremely low water absorption, excellent water vapor barrier properties, variable heat deflection properties, high rigidity, strength and hardness, very good blood compatibility, excellent biocompatibility, very good resistance to acids and alkalis, very good electrical insulating properties and very good processability/flowability. [0063] The polymeric material may include a cyclic olefin. The polymeric material may include cyclic olefin copolymer (COC). The polymeric material may include cyclic olefin polymers (COP).

[0064] The polymeric material may include a homopolymer or copolymer that includes polyethylene, polypropylene or both. The polymeric material may include a cyclic moiety. The polymeric material may include a polyester. The polymeric material may include polyester terephthalates or polyethylene terephthalate. The polymeric material may include polycarbonates. The polymeric material may include poly(methyl methacrylate).

[0065] The polymeric material may be processed in any suitable manner. For example, the polymeric material may be processed by methods for thermoplastics such as injection molding, extrusion, blow molding and injection blow molding.

[0066] The polymeric material may have excellent water vapor barrier properties. The polymeric material may have a low moisture vapor transmission rate (MVTR) or water vapor transmission rate (VWTR). The moisture vapor transmission rate may be determined according conventional testing methods such as DIN 53 122. The polymeric material may have a moisture vapor transmission rate of about 0.023 g ^■ mm/ m^{2 ■} d to about 0.045 g ^■ mm/ m^{2 ■} d at 23 °C and 85% relative humidity, as measured by DIN 53 122. The polymeric material may have a moisture vapor transmission rate of about 0.023 g ^■ mm/ m^{2 ■} d at 23 °C and 85% relative humidity. The polymeric material may have a moisture vapor transmission rate of less than about 0.023 g ^■ mm/ m^{2 ■} d at 23 °C and 85% relative humidity. The present teachings provide a blood collection tube which substantially prevents a reagent(s) from losing moisture over time. Moisture loss can cause the stabilizing reagent to become concentrated, which can potentially damage the integrity of all cells, including red blood cell lysis, producing excessive hemolysis.

[0067] The polymeric material may have high purity. The polymeric material may have low extractables and leachables. The polymeric material may be free of detectable residual metals. The polymeric material may be free of one or more of the following: phthalates, bisphenol A, heavy metals, halogens, fluorochemicals or fluoropolymers and vinyl chloride polymer or monomer. [0068] The polymeric material may allow for significant light transmission. The light transmission may be determined by ISO 13468-2. The polymeric material may have a light transmission of at least about 90%. The polymeric material may have a light transmission of greater than 90%.

[0069] The polymeric material may have a relatively low birefringence. Per ASTM D4093-95, the polymeric material may have a birefringence of about 1 nm to about 20 nm. The polymeric material may have a birefringence of less than 20 nm according to ASTM D4093-95. The polymeric material may have a birefringence of less than 15 nm according to ASTM D4093-95. The polymeric material may have a birefringence of less than 10 nm according to ASTM D4093-95. The polymeric material may have a birefringence of less than 5 nm according to ASTM D4093-95. The polymeric material may have a birefringence of less than about 2 nm according to ASTM D4093-95.

[0070] The polymeric material may be chemically resistant. For example, the polymeric material may have very good resistance to acid and alkalis. For example, the polymeric material may be resistant to one or more of the following: methanol, ethanol, isopropanol, acetone and butanone.

[0071] The polymeric material may be thermally stable. The coating may be thermally stable. The polymeric material and coating may each have coefficients of thermal expansion that are within about twenty percent of each other. The blood sample collection tube may be suitable for steam sterilization. The blood sample collection tube may be suitable for gamma sterilization.

[0072] The polymeric material may be an amorphous substance. Per ASTM D3417-99, for determination of heat of fusion and heat of crystallization of polymers by differential scanning calorimetry, the heat of fusion may be less than about 20 J/g. The heat of fusion as measured by differential scanning calorimetry (DSC) may be less than about 10 J/g. The heat of fusion as measured by differential scanning calorimetry (DSC) may be about less than 5 J/g. The heat of fusion as measured by differential scanning calorimetry (DSC) may be less than about 2 J/g.

[0073] The polymeric material may have a glass transition temperature of about 78 °C to about 136 °C as measured by differential scanning calorimetry (DSC). Preferably, the storage temperature range of the sample collection tube is from about 16 °C to about 35 °C. Preferably, the storage humidity of the blood sample collection tube is about 22% to about 79% relative humidity (RH).

[0074] The polymeric material may have a relatively low mold shrinkage. Thus, it is possible to use the material to help avoid distortion of geometry upon removal from a mold. The polymeric material may have a mold shrinkage of about 0.1 % to about 0.7%. The polymeric material may have a mold shrinkage of less than about 0.1 %.

[0075] The polymeric material may have a high strength and rigidity. The tensile strength and tensile modulus may be determined according to ISO 527 part 1 and 2. The polymeric material may have a tensile strength of about 46 MPa to about 63 MPa, as measured by ISO 527, parts 1 and 2. The polymeric material may have a tensile strength of about 60 MPa to about 63 MPa, as measured by ISO 527, parts 1 and 2. The polymeric material may have a tensile modulus of about 2600 MPa to about 3200 MPa, as measured by ISO 527, parts 1 and 2. The polymeric material may have a tensile modulus of about 2300 MPa to about 2600 MPa, as measured by ISO 527, parts 1 and 2. The impact strength may be determined according to ISO 1791 ell. The polymeric material may have an impact strength (Charpy Impact Unnotched) of about 13 kJ/m2 to about 20 kJ/m2, as measured by ISO 179/1 eU. The polymeric material may have an impact strength (Charpy Impact Unnotched) of about 20 kJ/m2, as measured by ISO 179/1 eU.

[0076] The sample collection tube may include a coating. The sample collection tube may be uncoated. The sample collection tube may be a blood collection tube with a coating on the inner wall of the tube. The coating may include one or more layers. The coating may be generally transparent. A layer of the coating may have a thickness of about 25 nm to about 1000 nm. A layer of the coating may have a thickness of about 50 nm. A layer of the coating may have a thickness of about 100 nm. The coating may have an average thickness throughout the tube of about 100 nm to about 500 nm.

[0077] The coating may serve one or more functions. The coating may contribute to the chemical stability of the polymeric blood collection tube. The coating may contribute to the mechanical stability of the polymeric blood collection tube.

[0078] The coating of the blood collection tube may be such that it alters one or more characteristics of the interior surface of the blood collection tube. For example, the coating may alter the surface energy of the inner walls of the tube. The coating may assist in preventing adherence of nucleic acids to the walls of the tube. The coating may assist in retaining a stopper in the tube when exposed to variable air pressures such as those encountered during transport by air.

[0079] The coating may include multiple layers. The coating may include one or more of the following layers: an adhesion layer, a barrier layer, a protective layer. Each layer may include different materials relative to an adjoining layer. Each layer may include the same or similar materials. The coating may include a silicon containing material. For example, the coating may include silicon oxide or silica. The coating may include one or more layers of silicon oxide. The coating may be free of metal oxide additives.

[0080] The coating may include a first layer, a second layer and a third layer. The first layer may be an adhesion layer deposited onto the inner wall of the polymeric sample collection tube. The second layer may be a barrier layer deposited onto the inner wall of the tube on top of the adhesion layer. The third layer may be a protective layer deposited onto the barrier layer. Thus, the barrier layer may be in between the adhesion layer and the protective layer.

[0081] The coating, including one or more layers, may be deposited on the inner wall of the tube via any suitable mechanism. For example the coating may be deposited onto the polymeric substrate by a plasma, a spray and/or a sputter deposition method. It may be applied by a vapor deposition method. It may be applied by a chemical deposition method. It may be applied by a physical deposition method. By way of one example, it may be applied by plasma enhanced chemical vapor deposition (PECVD).

[0082] In one example, the coating including one or more layers is deposited onto the polymeric substrate by plasma deposition. The plasma deposition method may be plasma-enhanced chemical vapor deposition. The plasma process may utilize monomers and gases. The plasma-enhanced chemical vapor deposition may include one or more of the following: an organosilicon precursor, oxygen, one or more noble gases. For example, the organosilicon precursor may include hexamethyldisiloxane (HMDSO), trimethylsilane (TriMS), tetramethylsilane (TetraMS), tetramethyldisiloxane (TMDSO), octamethylcyclotetrasiloxane (OMCTS) or a combination thereof. The blood collection tube may be vacuum sealed in an enclosure. The enclosure may include a gas inlet tube through which one or more gases and/or monomers are introduced into the enclosure. A plasma may be established inside the blood collection tube by radio frequency energy excitation. The plasma may deposit a uniform coating onto the interior surface of the blood collection tube.

[0083] The polymeric material of the blood collection tube is such that it is compatible with an adhesive backed label. The tube may include an adhesive-backed label adhered to an outer wall of the tube, and the adhesive may be an acrylic based adhesive. The adhesive backed label may resist peeling from the outer wall of the tube and/or outer portion of the stopper.

[0084] The blood collection tube of the present teachings may include a reagent. The tube may include a reagent with one or more ingredients. The ratio of one or more ingredients of the reagent may be about 5:1 to about 15: 1 (e.g. 10:1 ). The reagent may be solid. The reagent may be substantially solid. The reagent may be a liquid. The reagent may be a gel. The reagent may be a film.

[0085] The reagent may include an aqueous substance. The reagent may include one or more agents in solution. Suitable solvents may include water, saline, dimethy!su!foxide, alcohol and any mixture thereof

[0086] The reagent may include a preservative. The reagent may include an anticoagulant. The reagent may include a preservative and an anticoagulant. The tube may include a preservative and optionally an anticoagulant. The reagent may include a preservative, an anticoagulant and one or more other agents. For example, the reagent may include one or more aldehyde reaction agents. For example, the aldehyde reaction agent may be selected from one or any combination of tris, glycine, or a derivative (e.g., a salt and/or an ester) of either or both. The reagent may include one or more preservative agents, one or more enzyme inhibitors, one or more metabolic inhibitors, or any combination thereof. The reagent may include glycine. For example, the blood collection tube may include the preservative imidazolidinyl urea (IDU), the anticoagulant EDTA and glycine. The tube may include a reagent with one or more ingredients and a ratio of the one or more ingredients may be about 5: 1 to about 15: 1 . The tube may include imidazolidinyl urea (IDU) and glycine and a ratio of imidazolidinyl urea (IDU) to glycine may be about 10:1 . [0087] The aldehyde reaction agent may be present in a ratio (by weight) relative to the preservative composition of about 1 :20 to about 1 : 1 . The aldehyde reaction agent may be present in a ratio (by weight) relative to the anticoagulant of about 1 :25 to about 5: 1 . The preservative composition may be present in a ratio (by weight) relative to the anticoagulant of about 1 : 10 to about 15:1 .

[0088] The reagent may include one or more amines that may be selected from one or more of tryptophan, tyrosine, phenylalanine, glycine, ornithine and S- adenosylmethionine, aspartate, glutamine, alanine, arginine, cysteine, glutamic acid, glutamine, histidine, leucine, lysine, proline, serine, threonine, or combinations thereof. The one or more amines may be chosen based upon their reactive capabilities. As one non-limiting example, it is possible that the one or more amines be aldehyde reaction agents. For example, the aldehyde reaction agent may be selected from one or any combination of tris, lysine, glycine, urea, or a derivative (e.g., a salt and/or an ester) of either or both. The aldehyde reaction agent may be selected to react with any free formaldehyde that may be present either prior to or post sample collection. The concentration of the amines in the reagent (prior to blood draw) may be from about 0.25% to about 1 .5%. The concentration of the one or more amines may be from about 0.3% to about 0.8%. The concentration of the one or more amines may be about 0.4% to about 0.7%.

[0089] The reagent may include one or more anticoagulants or chelating agents which may be selected from the group consisting of ethylene diamine tetra acetic acid (EDTA) and its salts, ethylene glycol tetra acetic acid (EGTA) and its salts, hirudin, heparin, citric acid, salts of citric acid, oxalic acid, salts of oxalic acid, acid citrate dextrose (ACD), citrate, citrate-theophylline-adenosine-dipuridamole (CTAD), citrate- pyridoxalphosphate-tris, heparin-p-hydroxy-ethyl-theophylline, polyanethol sulfonate, sodium fluoride, sodium heparin, thrombin and PPACK (D-phenylalanyl-L-prolyl-L- arginine chloromethyl ketone and any combination thereof. Thus the concentration of the anticoagulant in the reagent (prior to blood draw) may be from about 2.5% to about 10%. The concentration of the anticoagulants may be from about 3% to about 8%. The concentration of the anticoagulants may be about 4% to about 7%. [0090] The reagent may include one or more polysaccharides, which may be selected from starch, cellulose, glycogen, or combinations thereof. The one or more polysaccharides may act as erythrocyte protectant agents. The one or more polysaccharides may assist in stabilizing erythrocyte membranes, such that cell lysis is slowed, minimized, substantially prevented or some combination thereof. The concentration of the polysaccharides in the reagent (prior to blood draw) may be from about 0.001 % to about 5.0%. The concentration of the polysaccharides in the reagent may be from about 0.02% to about 3.0%. The concentration may be about 0.5% to about 2.0%.

[0091] The reagent may include one or more transcription inhibitors, which may be selected to promote stability of one or more components within a sample. The one or more transcription inhibitors may be selected from actinomycin D, a-amanitin, triptolide, 5,6-dichloro-1 -p-D-ribofuranosylbenzimidazole (DRB), flavopiridol, or any combination thereof. The concentration of the one or more transcription inhibitors may be about 0.5 μΜ to about 500 μΜ.

[0092] The reagent may include one or more enzyme inhibitors. The one or more enzyme inhibitors may be selected from the group consisting of: diethyl pyrocarbonate, ethanol, aurintricarboxylic acid (ATA), glyceraldehydes, sodium fluoride, ethylenediamine tetraacetic acid (EDTA), formamide, vanadyl-ribonucleoside complexes, macaloid, heparin, hydroxylamine-oxygen-cupric ion, bentonite, ammonium sulfate, dithiothreitol (DTT), beta-mercaptoethanol, cysteine, dithioerythritol, tris(2-carboxyethyl)phosphene hydrochloride, a divalent cation such as Mg+2, Mn+2, Zn+2, Fe+2, Ca+2, Cu+2 and any combination thereof.

[0093] The reagent may include one or more metabolic inhibitors. The one or more metabolic inhibitors may be selected from the group consisting of: glyceraldehyde, dihydroxyacetone phosphate, glyceraldehyde 3-phosphate, 1 ,3-bisphosphoglycerate, 3- phosphoglycerate, 2-phosphoglycerate, phosphoenolpyruvate, pyruvate and glycerate dihydroxyacetate, sodium fluoride, K2C2O4 and any combination thereof.

[0094] The reagent may include one or more protease inhibitors. The one or more protease inhibitors may be selected from the group consisting of: antipain, aprotinin, chymostatin, elastatinal, phenylmethylsulfonyl fluoride (PMSF), APMSF, TLCK, TPCK, leupeptin, soybean trypsin inhibitor, indoleacetic acid (IAA), E-64, pepstatin, VdLPFFVdL, EDTA, 1 ,10-phenanthroline, phosphoramodon, amastatin, bestatin, diprotin A, diprotin B, alpha-2-macroglobulin, lima bean trypsin inhibitor, pancreatic protease inhibitor, egg white ovostatin egg white cystatin, and any combination thereof. The protective agent may include a phosphatase inhibitor selected from the group consisting of: calyculin A, nodularin, NIPP-1 , microcystin LR, tautomycin, okadaic acid, cantharidin, microcystin LR, okadaic acid, fostriecin, tautomycin, cantharidin, endothall, nodularin, cyclosporin A, FK 506/immunophilin complexes, cypermethrin, deltamethrin, fenvalerate, bpV(phen), dephostatin, mpV(pic) DMHV, sodium orthovanadate and any combination thereof.

[0095] The reagent may include one or more nuclease inhibitors. The one or more nuclease inhibitors may be selected from the group consisting of: diethyl pyrocarbonate, ethanol, aurintricarboxylic acid (ATA), formamide, vanadyl-ribonucleoside complexes, macaloid, ethylenediamine tetraacetic acid (EDTA), proteinase K, heparin, hydroxylamine-oxygen-cupric ion, bentonite, ammonium sulfate, dithiothreitol (DTT), beta-mercaptoethanol, cysteine, dithioerythritol, tris(2-carboxyethyl) phosphene hydrochloride, or a divalent cation such as Mg+2, Mn+2, Zn+2, Fe+2, Ca+2, Cu+2 and any combination thereof.

[0096] The reagent may include one or more phosphatase inhibitors. The one or more phosphatase inhibitors may be selected from the group consisting of: calyculin A, nodularin, NIPP-1 , microcystin LR, tautomycin, okadaic acid, cantharidin, imidazole, microcystin LR, okadaic acid, fostriecin, tautomycin, cantharidin, endothall, nodularin, cyclosporin A, FK 506/immunophilin complexes, cypermethrin, deltamethrin, fenvalerate, bpV(phen), dephostatin, mpV(pic) DMHV, sodium orthovanadate and any combinations thereof.

[0097] The reagent may include bicyclic oxazolidines, DMDM hydantoin, sodium hydroxymethylglycinate, hexamethylenetetramine chloroallyl chloride, biocides, a water- soluble zinc salt or any combination thereof. The reagent may include a polyacrylic acid or a suitable acid having a pH ranging from about one to about seven. The reagent may include amines, amino acids, alkyl amines, polyamines, primary amines, secondary amines, ammonium salts, or any combination thereof. The reagent may include one or more primary amines. The reagent may include one or more amides (e.g. butanamide). The reagent may include one more apoptosis inhibitors. The reagent may include one or more caspase inhibitors.

[0098] The reagent may include one or more polymer ingredients. The polymers may include but are not limited to the following: polyvinylpyrrolidone (PVP), polyethylene glycol (PEG) and cyclodextrin.

[0099] The reagents described herein may also include additional components, including one or more of the following: Doxycycline, Polyethylene Glycol, Sulfasalazine, Polyvinylpyrrolidone, Curcumin, Magnesium Gluconate, Homocysteine, Methyl Cellulose (MC), 6-Aminocaproic acid, Ethyl Cellulose, Aprotinin, Hydroxyethyl Cellulose, Doxycycline, Hydroxypropyl Cellulose, Minocycline HCI, Dextrin, Nicotinamide, Dextran, Chitosan, Polyethylene Oxide, Lysine, Poly Ethyl Oxazoline, Glyceraldehyde, Ficolls, Phytic Acid, a-Cyclodextrin, b-Sitoserol, β-Cyclodextrin, C-AMP, Y-Cyclodextrin, Poly Lysine, Gelatins, Biochanin A, Sugars (e.g., sucrose, mannitol, lactose, trehalose), Sulfasalazine, Hydroxypropyl Methyl Cellulose, Demeclocycline, Hydroxyethyl Methyl Cellulose, Chlortetracycline, Oxytetracycline, Cyclohexamide, Rifampicin, Soy Milk, soybean based protease inhibitor, Suramin, N-Butyric Acid, Penicillamine, N-Acetyl Cysteine, Benzamidine, AEBSF, Alpha-2 Macroglobulin, or combinations thereof.

[00100] The reagents may include one or more cell permeabilizing agents. The one or more cell permeabilizing agents may be selected from the group consisting of: DMSO (dimethyl sulfoxide), ethylene glycol, polyethylene glycol, glycerin, Cellosolves (ethylene glycol dimethyl ether) (phenoxyethanol), Triton X 100, Triton X 705 (non-ionic detergents), 1 -methyl-2-pyrrolidinone, Tween 20, Tween 40 (non-ionic), Brij 35 (detergent), polyoxyethylene ether (Polyox), sodium cholate, ethylene oxide polymers, monensin, monactin, pentachlorophenol, 2,4 dinitrophenol, saponin, SDS (sodium dodecyl sulfate) and combinations thereof.

[00101] The reagent may include proteins such as: biotin, albumins (egg, bovine), gelatin, and similar such compounds. The reagent may include RNAse inhibitors such as: human placenta derived RNAse inhibitor, and similar such compounds. The reagent may include nucleic acid stabilizers such as: guanidinium hydrochloride, polycations such as polyethylenimine, and similar such compounds. The reagent may include amino acids/polypeptides such as: glutamic acid, glycine, aspartic acid, and similar such compounds. The reagent may include fixatives such as: aldehydes (formaldehyde and glutaraldehyde), alcohols (ethanol, methanol), and similar such compounds. The reagent may include anticoagulants such as: EDTA (Ethylene Diamine Tetra acetic acid), and similar such compounds. The reagent may include ACD (Acid Citrate Dextrose), Heparin, and similar such compounds. The reagent may include protease inhibitors such as: EDTA, PMSF (phenyl methyl sulfonyl fluoride), AEBSF (2-Aminoethyl benzene sulfonyl fluoride), and similar such compounds. The reagent may include antioxidants/reducing agents such as: Trolox, a-tocopherol, B-mercaptoethanol, and similar such compounds. The reagent may include nucleic acid dyes such as: DAPI (diamidino 2-phenylindole), propidium iodide, fluorescein diacetate, and similar such compounds. The reagent may include carbohydrates such as: sugars (sucrose), cellulose, and similar such compounds. It should be appreciated that the above specific listings of compounds may contain a measure of overlap, which recognizes the sometimes-overlapping function of certain specific compounds. One of skill in the art should understand and appreciate this aspect of the disclosure.

[00102] The reagent may be present in the tube as a liquid in an amount less than about 10% by volume of the tube but greater than about 0.1 % by volume. The reagent may be present in the tube as a liquid in an amount less than about 5% by volume of the tube but greater than about 0.1 % by volume of the tube. The reagent may be present in the tube in an amount less than about 3% by volume of the tube but greater than about 0.1 % by volume of the tube. It is possible that at any point in time (e.g., prior to blood draw, just following blood draw, 1 hour post blood draw, 12 hours post blood draw, 4 days post blood draw) the tube may contain a detectable amount of formaldehyde. Such detection may be facilitated by use of an acetylacetone test. Thus it is possible that the tube contains blood plasma, a preservative, an anticoagulant, and formaldehyde. It is further possible that the tube contain blood plasma, a preservative, an anticoagulant, formaldehyde, and a plurality of cell-free nucleic acids (e.g., cell-free DNA (which may be cell-free fetal DNA, cell free RNA (which may be cell-free mRNA), circulating tumor- derived nucleic acids, or nucleic acids of other origin).

[00103] The blood collection tube of may include a blood draw volume of about 4 ml to about 10 ml. The blood collection tube may include a reagent volume of about .20 ml to about .90 ml. The blood collection tube may include about .20 ml to about .30 ml of reagent for a blood draw volume of about 8 ml to about 10 ml. The blood collection tube may include about .60 ml to about .90 ml of reagent for a blood draw volume of about 4 ml to about 6 ml.

[00104] The reagent in the tube may include about 15% to about 70% by volume of the preservative composition prior to blood draw. The reagent in the tube may include about 15% to about 70% by volume of imidazolidinyl urea (IDU) prior to blood draw. The reagent in the tube include may include about 5% to about 20% by volume of anticoagulant prior to blood draw. The reagent in the tube may include about 5% to about 20% of anticoagulant prior to blood draw. The reagent in the tube may include about 5% to about 10% of anticoagulant prior to blood draw. The reagent in the tube may include about 1 % to about 10% of an agent in addition to the preservative and an anticoagulant prior to blood draw.

[00105] The reagent may include one or more preservative agents. The preservative may be selected from the group consisting of formaldehyde, diazolidinyl urea (DU), imidazolidinyl urea (IDU), dimethylol urea, 2-bromo-2-nitropropane-1 ,3-diol, 5- hydroxymethoxymethyl-1 -aza-3,7-dioxabicyclo (3.3.0)octane and 5-hydroxymethyl-1 - aza-3,7-dioxabicyclo (3.3.0)octane and 5-hydroxypoly [methyleneoxy]methyl-1 -aza-3,7- dioxabicyclo (3.3.0)octane, bicyclic oxazolidines, DMDM hydantoin, sodium hydroxymethylglycinate, hexamethylenetetramine chloroallyl chloride, biocides, a water- soluble zinc salt and any combination thereof. The one or more preservative agents may include a formaldehyde releaser such as one selected from the group consisting of: diazolidinyl urea, imidazolidinyl urea, dimethoylol-5,5-dimethylhydantoin, dimethylol urea, 2-bromo-2.-nitropropane-1 ,3-diol, oxazolidines, sodium hydroxymethyl glycinate, 5- hydroxymethoxymethyl-1 -1 aza-3,7-dioxabicyclo [3.3.0]octane, 5-hydroxymethyl-1 -1 aza- 3,7dioxabicyclo[3.3.0]octane, 5-hydroxypoly[methyleneoxy]methyl-1 -1 aza-

3,7dioxabicyclo[3.3.0]octane, quaternary adamantine and any combination thereof.

[00106] The reagent may contain less than about 20 parts per million (ppm) of an aldehyde per 100 μΙ of the reagent (prior to contact with a sample). The reagent may contain less than about 15 parts per million (ppm) of an aldehyde per 100 μΙ of the reagent. The reagent may contain less than about 10 parts per million (ppm) of an aldehyde per 100 μΙ of the reagent. The reagent may contain less than about 5 parts per million (ppm) of an aldehyde per 100 μΙ of the reagent. The reagent may contain at least about 0.1 parts per million (ppm) per 100 μΙ of the reagent. The reagent may contain at least about 0.5 parts per million (ppm) to about 15 ppm of an aldehyde per 100 μΙ of the reagent. The reagent may contain at least about 1 part per million (ppm) of an aldehyde per 100 μΙ of the reagent. The reagent may contain at least about 10 parts per million (ppm) of an aldehyde per 100 μΙ of the reagent. The reagent may contain at least about 100 parts per million (ppm) of an aldehyde per 100 μΙ of the reagent. The reagent may contain at least about 1000 parts per million (ppm) of an aldehyde per 100 μΙ of the reagent. The reagent may contain at least about 10,000 parts per million (ppm) of an aldehyde per 100 μΙ of the reagent. The reagent may contain less than 1 parts per million (ppm) an aldehyde per 100 μΙ of the reagent after contact with a sample. The aldehyde may include formaldehyde.

[00107] The reagent may include one or more aldehyde reaction agents. The reagent may include an amount of aldehyde reaction agent containing a primary amine for reacting with free formaldehyde present in the reagent to result in an aldehyde amount below 10,000 ppm per 100 μΙ of reagent prior to blood draw. The reagent may include an amount of aldehyde reaction agent, which aldehyde reaction agent is selected from one or any combination of tris, glycine, or a derivative (e.g., a salt and/or an ester) of either or both, for reacting with free formaldehyde present in the reagent to result in an aldehyde amount below 10,000 ppm per 100 μΙ of reagent prior to blood draw.

[00108] It will be appreciated that the above is by way of illustration only. Other ingredients may be employed in the sample collection tube disclosed herein, as desired, to achieve the desired resulting characteristics. Examples of other ingredients that may be employed include antibiotics, anesthetics, antihistamines, preservatives, surfactants, antioxidants, unconjugated bile acids, mold inhibitors, nucleic acids, pH adjusters, osmolarity adjusters, or any combination thereof.

[00109] General applications of the present teachings may be found in as described in U.S. Patent Application Nos. US20040137417, US20100184069, and U.S. Patent No. 8304187. In general, the blood collection tubes described herein may be filled with one or more reagents as described in U.S. Patent Application Nos. US20040137417, US 20100184069, and U.S. Patent No. 8304187.

[00110] The blood collection tube may be non-reactive to one or more ingredients. For example, the blood collection tube may be non-reactive to anticoagulants. The blood collection tube may be non-reactive to preservatives. The blood collection tube may be non-reactive to the collected blood sample. The blood collection tube may be non- reactive to the combination of the one or more ingredients and the collected blood sample. Per ISO 10993, the blood collection tubes of the present teachings may be compliant with testing standards which demonstrate the biocompatibility of medical devices such as blood collection tubes. The reagent may include or the tube may be adapted to receive one or more materials for enabling isolation of a cellular or cell-free component of a blood sample. Such materials may include a protease or amino acid which may include but is not limited to proteinase K.

[00111] The tube may be suitable for storing a blood sample for a period of at least about 7 days. The tube may be suitable for storing a blood sample for a period of at least about 14 days. The tube may be suitable for storing a blood sample for a period of at least about 30 days. The tube may be suitable for storing a blood sample for a period of at least about 60 days. The tube may be suitable for storing a blood sample for a period of at least about 90 days.

[00112] It is believed that a blood sample stored in the blood collection tube of the present teachings will result in a substantial increase in the amount of recoverable nucleic acid from the blood sample (as compared to a sample collected in a standard tube and/or a polymeric tube with no coating). A blood sample stored in the tube having a silicon containing coating may result in a substantial increase in the amount of recoverable nucleic acid or rare cells as compared with a polymeric tube of the same polymer but with no coating and the substantial increase may be at least about 10%. A blood sample stored in the tube having a silicon-containing coating may result in a substantial increase in the amount of recoverable nucleic acid or rare cells as compared with a polymeric tube of the same polymer but with no coating and the substantial increase may be at least about 20%. A blood sample stored in the tube having a silicon-containing coating may result in a substantial increase in the amount of recoverable nucleic acid or rare cells as compared with a polymeric tube with no coating and the substantial increase may be at least about 30%.

[00113] FIG.1 depicts an illustrative example of the blood collection tube assembly 300 of the present teachings. The blood collection tube 100 is cylindrical in shape. The tube 100 includes a stopper 200 inserted in the opening 20 of the tube 100. There is an interface 150 between the outer wall of the stopper 200 and the inner wall of the tube 100.

[00114] FIGS. 2(a)-(c) depict an illustrative example of a 4 ml blood collection tube. The tube 100 includes an opening 20. The opening 20 of the tube 100 includes an annular rim 30. The tube 100 includes a base portion 40 opposing the opening 20. The tube 100 tapers in diameter to the base 40. The base 40 includes a recessed dimple 50. FIG. 2(b) illustrates a close-up view of portion B of the tube 100 of FIG. 2(a) depicting the upper portion of the tube 100 terminating at an open end and including an annular rim 30. The annular rim 30 is enlarged relative to the coextensive sidewall of the tube 100. FIG. 2(c) illustrates a close-up view of portion C of the tube 100 of FIG. 2(a) depicting the lower portion of the tube 100 including a base 40 and recessed dimple 50.

[00115] FIGS. 3(a)-(c) depict an illustrative example of a 10 ml blood collection tube. The tube 100 includes an opening 20. The opening 20 of the tube 100 includes an annular rim 30. The tube 100 includes a base portion 40 opposing the opening 20. The tube 100 tapers in diameter to the base 40. The base 40 includes a recessed dimple 50. FIG. 3(b) illustrates a close-up view of portion B of the tube 100 of FIG. 3(a) depicting the upper portion of the tube 100 terminating at an open end and including an annular rim 30. The annular rim 30 is enlarged relative to the coextensive sidewall of the tube 100. FIG. 3(c) illustrates a close-up view of portion C of the tube 100 of FIG. 3(a) depicting the lower portion of the tube 100 including a base 40 and recessed dimple 50.

[00116] Draw Volume Verification

[00117] FIG. 4(a) depicts the results of draw volume verification for twenty 9 ml blood collection tubes of the present teachings. The water draw volume was within 9 ml_ ± 10% for each tube. The tare weight of the twenty 9 ml tubes was recorded. Water was vacuum drawn into each tube and weighed post draw. The weight to milliliter conversion for this study is 1 .0 g = 1 .0 ml_. The water draw volume was to be within 9 ml_ ± 10% for each tube in accordance with ISO 6710. The results demonstrate that the water draw volume of each polymeric coated tubes including reagent was within the specification, as shown in FIG. 4(a), with an average of 9.45 mL (StDev: 0.018 mL). (The tubes are manufactured such that the minimum volume for a properly drawn sample at the beginning of product dating is 9.0 mL.)

[00118] FIG. 4(b) depicts the results of draw volume verification for twenty 4 ml blood collection tubes of the present teachings. The water draw volume was within 4 mL ± 10% for each tube. The tare weight of the twenty 4 ml tubes was recorded. Water was vacuum drawn into each tube and weighed post draw. The weight to milliliter conversion for this study is 1 .0 g = 1 .0 mL. The water draw volume was to be within 4 mL ± 10% for each tube in accordance with ISO 6710. The results demonstrate that the water draw volume of each polymeric coated tubes including reagent was within the specification, as shown in FIG. 4(b), with an average of 3.959 mL (StDev: 0.028 mL). (The tubes are manufactured such that the minimum volume for a properly drawn sample at the beginning of product dating is 4.0 mL.)

[00119] Plasma Recovery

[00120] The present teachings demonstrate there is no statistically significant difference between plasma recovery from glass blood collection tubes and the blood collection tubes of the present teachings. Plasma recovery was tested utilizing 10 mL glass blood collection tubes and 9 mL blood collection tubes of the present teachings. In protocol #1 , whole blood was centrifuged at 300 x g for 20 minutes at room temperature to separate plasma. The plasma recovery was 4.5 mL for a 10 mL glass tube and 4.1 mL for a 9.0 mL blood collection tube of the present teachings (< 10% difference). In protocol # 2, whole blood was centrifuged at 1 ,600 x g for 10 minutes at room temperature to separate plasma. The plasma recovery was 5.5 mL for a 10 mL glass tube and 5.0 mL for a 9.0 mL blood collection tube of the present teachings (< 10% difference). (The blood collection tubes of the present teachings will see approximately 10% less plasma due to the draw volume difference from 10 mL to 9 mL.)

[00121] Performance Testing - Cell-Free Plasma DNA

[00122] FIGS. 5-7 depict cell-free DNA genome equivalents per 1 mL of plasma for a storage temperature study assessing glass blood collection tubes and the blood collection tubes of the present teachings. FIG. 8 depicts cell-free DNA genome equivalents per 1 ml_ of plasma for a shipping and handling study assessing glass blood collection tubes and the blood collection tubes of the present teachings. The present teachings demonstrate there is no statistically significant difference between the performance (i.e., stabilization attributes) of a cell-free nucleic acid reagent, such as that utilized in Cell-Free DNA BCT® by Streck (Omaha, Nebraska), in glass evacuated blood collection tubes and the evacuated blood collection tubes of the present teachings as it relates to cellular DNA contamination of cell-free DNA (cfDNA) in plasma during whole blood sample storage, shipping and handling.

[00123] The impact of storage temperature and shipping and handling was assessed. Both blood collection tube types were sterilized via gamma irradiation to a sterility assurance level (SAL) of 10-3, per ISO 1 1 137. The lone experimental variable was the material the vials are made of. The rubber stopper/septum and the Cell-Free DNA BCT® reagent are the same in both package configurations. Donors were from both sexes and presumed to be healthy. Blood was collected from donors according to the protocol in the instructions for use for Cell-Free DNA BCT® in both glass evacuated blood collection tubes and the evacuated blood collection tubes of the present teachings. Both blood collection tube types were mixed immediately after blood draw by inverting ten times per the instructions for use.

[00124] To assess the impact of storage temperature, five donors had blood drawn into three glass evacuated blood collection tubes and three blood collection tubes of the present teachings for each temperature (6 ± 2 °C, 18-25 °C, and 37 ± 2 °C). After collection, three glass and three polymeric coated blood collection tubes from each donor were stored at 6 ± 2 °C. The plasma was separated by centrifugation and the plasma cfDNA was isolated and quantified from one tube of each type on day 1 , one tube of each type on day 7, and one tube of each type on day 14 (as described below). This study was repeated with new donors at storage temperatures of 18-25 °C and 37 ± 2 °C.

[00125] To assess the impact of shipping and handling, five donors had blood drawn into three glass evacuated blood collection tubes and three blood collection tubes of the present teachings. One polymeric coated tube and one glass tube per donor was packed in an insulated box equipped with a temperature monitoring device (Omega Nomad, Omega Engineering, Stamford, CT) and shipped from Omaha, NE, round trip via overnight air freight to a laboratory out of state (elapsed time 48 hours). Upon return, the plasma was separated and the plasma cfDNA was isolated and quantified (day 2). Two polymeric coated and two glass tubes per donor that were not shipped were maintained at room temperature (18-25 °C). The plasma was separated and the plasma cfDNA was isolated and quantified from one tube of each type on days 0 and 2.

[00126] The blood plasma separation process included a 2-step process. First, blood was mixed and centrifuged in each blood collection tube at 18-25 °C at an initial low speed centrifugation at 300 x g for 20 minutes. Second, the plasma layer was carefully removed without disturbing the buffy coat, transferred to a new vial, and centrifuged at 18-25 °C at 5,000 x g for 10 minutes to remove residual cells. Plasma was stored at -80 °C until cell-free DNA isolation or processed immediately. This process was performed at each testing interval for both the storage temperature and shipping and handling studies.

[00127] Cell-free DNA was isolated from plasma. Extraction of cell-free DNA can be accomplished using most commercially available kits. Specific to this evaluation, the QIAamp® Circulating Nucleic Acid Kit (Qiagen, Santa Clarita, CA) was used for extraction of cell-free DNA from the plasma complement. The manufacturer's recommended protocol was modified to increase the duration of the Proteinase K treatment from 30 minutes to 1 hour at 60 °C per the instructions for use protocol. Isolated cfDNA was stored at -80 °C until analysis by Droplet Digital Polymerase Chain Reaction (ddPCR).

[00128] Cell-free DNA was quantified by ddPCR. The following primers and probe for this study were designed for the quantification of the human β-Actin housekeeping gene. Forward primer: 5'-GGT GTT TGT CTC TCT GAC TAG G-3'; reverse primer: 5'- CAC ACG AGC CAG TGT TAG TA-3'; probe: 5'-HEX-TGT CTG AGA CAG TGT TGT GGG TGT-BHQ-3'. The primers and probe were synthesized by Integrated DNA Technologies (Coralville, IA). A PCR master mix, 2X ddPCR™ Supermix for Probes, was purchased from Bio-Rad Laboratories (Hercules, CA). Final concentrations of primers and probe in PCR reactions were 900 nM and 250 nM, respectively, in a final volume of 20 μΙ. A Bio-Rad QX100 Droplet Digital™ PCR System was used. Thermal cycling was performed with a Bio-Rad C1000 Touch Thermal Cycler. The following PCR conditions were used: 10 minutes at 98 °C; 40 cycles of 30 seconds at 95 °C, 30 seconds at 54 °C, and 30 seconds at 72 °C. A final extension of 10 min at 72 °C was followed by a heating step for 10 minutes at 98 °C to inactivate the polymerase. Data analysis was performed using Bio-Rad QuantaSoft software version 1 .3.2. Duplicate PCR reactions were set-up for each testing interval and cfDNA sample isolated. Results were reported in Genome Equivalent/mL of Plasma (GE/mL). GE/mL = genome equivalents/milliliter- One genome equivalent is defined as the amount of target sequence (i.e., β-Actin for the purposes of this validation) present in one diploid cell. The acceptance criteria for all polymeric coated vs. glass equivalency studies was a genome equivalent/mL fold increase < 4.

[00129] The results indicate that acceptance criteria for all tests and conditions evaluated was met as the average fold increase of genome equivalent/mL of plasma of the blood collection tube of the present teachings was < 4. The GE/mL results of the storage temperature and shipping and handling studies are shown in FIGS 5-8. Box plots show the median (line inside the box) and 75th and 25th percentiles (limits of the box). The upper and lower error bars indicate the 90th and 10th percentiles, respectively. The upper most and lower most dots indicate the maximum and minimum values.

[00130] Performance Testing - Circulating Tumor Cells

[00131] The present teachings demonstrate there is no statistically significant difference between the performance of a cell-free nucleic acid reagent, such as that utilized in Cell-Free DNA BCT® by Streck (Omaha, Nebraska), in glass evacuated blood collection tubes and the evacuated blood collection tubes of the present teachings as it relates to the stabilization of circulating tumor cells (CTC) during whole blood sample storage at 15-30 °C. The effective stabilization of CTCs with blood drawn into glass Cell- Free DNA BCT® has been shown. (Qin et al. Stabilization of circulating tumor cells in blood using a collection device with a preservative reagent. Cancer Cell International 2014, 14:23.)

[00132] Three donors had blood drawn into two polymeric coated tubes including reagent. Blood collection tubes were mixed immediately after blood draw by inverting ten times per the instructions per use for Cell-Free DNA BCT®. All samples were spiked with a known number of Michigan Cancer Foundation breast cancer cells (MCF-7) and immediately mixed by inverting ten times. Samples were shipped overnight to a laboratory out of state. Samples were analyzed on day 7 on the Veridex CellSearch™ (Raritan, New Jersey) system in order to quantify the recovery rate of MCF-7 cells stabilized in polymeric coated tubes including the reagent utilized in Cell-Free DNA BCT®. Mean CTC % recovery of the polymeric coated tubes including Cell-Free DNA BCT® reagent was to be within range of previously published CTC % recovery data of 52-60% by Qin et al.

[00133] The results indicate mean CTC % recovery of the polymeric coated tubes including Cell-Free DNA BCT® reagent was within acceptable range over a period of 7 days at 15-30 °C. Data shown in Table 1 supports the instructions for use claim that the formaldehyde-free preservative contained in Cell-Free DNA BCT® stabilizes circulating tumor cells in whole blood for up to 4 days at temperatures between 15-30 °C.

[00134] Table 1. Mean CTC recovery at day 7 for polymeric coated tube with reagent.

[00135] Performance Testing - Flow Cytometry

[00136] The present teachings demonstrate there is no statistically significant difference between the performance (i.e., stabilization attributes) of a immunophenotyping preservative reagent, such as that utilized in Cyto-Chex BCT® by Streck (Omaha, Nebraska), in glass evacuated blood collection tubes and the evacuated blood collection tubes of the present teachings as it relates to immunophenotyping of white blood cells during whole blood sample storage, shipping and handling.

[00137] The impact of storage temperature and shipping and handling was assessed. Both blood collection tube types were sterilized via gamma irradiation to a sterility assurance level (SAL) of 10-3, per ISO 1 1 137. The lone experimental variable was the material the vials are made of. The rubber stopper/septum and the Cyto-Chex BCT® reagent are the same in both package configurations. Donors were from both sexes and presumed to be healthy. Blood was collected from donors according to the protocol in the instructions for use for Cyto-Chex BCT® in both glass evacuated blood collection tubes and the evacuated blood collection tubes of the present teachings. Both blood collection tube types were mixed immediately after blood draw by inverting eight to ten times per the instructions for use.

[00138] To assess the impact of storage temperature, three donors had blood drawn into three glass evacuated blood collection tubes, three blood collection tubes of the present teachings, and one EDTA tube. After collection, one glass and one blood collection tube of the present teachings from each donor were stored at each of the three temperatures (18 °C to 22 °C, 30 °C and 37 ^°C). The EDTA tube sample was analyzed to establish a baseline of CD marker recovery for the HIV panel (CD3, CD4, CD8, CD19, CD16/56, and CD45).

[00139] The glass tube and the blood collection tube of the present teachings stored at 37 °C were tested 48 hours post blood draw for CD8. The same two tubes were tested 72 hours post blood draw for CD3 and CD4. The glass tube and the blood collection tube of the present teachings stored at 30 °C were tested 7 days post blood draw for the HIV panel of markers (CD3, CD4, CD8, CD19, CD16/56, and CD45). The glass tube and the blood collection tube of the present teachings stored at 18 °C to 22 °C were tested 7 days and 14 days post blood draw for the HIV panel of markers (CD3, CD4, CD8, CD19, CD16/56, and CD45).

[00140] To assess the impact of shipping and handling, three donors had blood drawn into three glass evacuated blood collection tubes, three blood collection tubes of the present teachings, and one EDTA tube. One blood collection tube of the present teachings and one glass tube per donor remained on site and were placed at room temperature (18 °C to 22 °C). One blood collection tube of the present teachings and one glass tube per donor were packed in an insulated box equipped with a temperature monitoring device (Omega Nomad, Omega Engineering, Stamford, CT) and shipped from Omaha, NE, round trip via overnight air freight to a laboratory out of state (elapsed time 48 hours). Upon return receipt, the blood collection tube of the present teachings and the glass tube were placed at room temperature (18 °C to 22 °C). All four tubes were tested 7 days and 14 days post blood draw for the HIV panel of markers (CD3, CD4, CD8, CD19, CD16/56, and CD45). [00141] For all studies and conditions in the protocol, flow cytometry testing was performed on the BD FACSCalibur™ with Multiset™ software V3.0.2 using 4-color BD Multitest reagents and a lyse/no-wash sample preparation procedure.

[00142] The acceptance criteria is as follows: the % difference of CD marker recovery between EDTA tubes and the blood collection tubes of the present teachings must be < 20% for all studies and testing conditions.

[00143] The results indicate the % difference of all CD markers between EDTA tubes and the blood collection tubes of the present teachings at each testing condition was < 20%. The HIV panel of markers including CD3, CD4, CD8, CD16/56, CD19 and CD45 are stable in blood samples stored in the blood collection tubes of the present teachings for up to 14 days at 18 °C to 22 °C and up to 7 days at 30 °C. CD8 is stable for up to 48 hours at 37 °C. CD3 and CD4 are stable for up to 72 hours at 37 °C. Immunophenotypic analysis performed on peripheral blood samples stored in the blood collection tubes of the present teachings provides the same results as when performed on fresh specimens as shown in Tables 2-10.

[00144] Table 2. Percent difference of cell count recovery between day 7 glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for storage temperature study at 18 °C to 22 °C.

[00145] Table 3. Percent difference of cell count recovery between day 14 glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for storage temperature study at 18 °C to 22 °C. Polymeric Coated

EDTA Glass Tube

Tube

Target Data Initial Day 14 % Day 14 % Markers Points Cells/μΙ Cells/μΙ Difference Cells/μΙ Difference

CD3 12 11 11 1087 2.2% 1071 3.6%

CD4 12 766 748 2.2% 756 1.3%

CD8 12 329 309 6.1% 289 12.3% CD16/CD56 12 238 212 11.0% 227 4.6% CD19 12 127 133 4.3% 119 6.2% CD45 12 1522 1507 1.0% 1489 2.2%

[00146] Table 4. Percent difference of cell count recovery between day 7 glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for storage temperature study at 30 °C.

[00147] Table 5. Percent difference of cell count recovery between 48-hour glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for storage temperature study at 37 °C.

[00148] Table 6. Percent difference of cell count recovery between 72-hour glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for storage temperature study at 37 °C. Polymeric Coated

EDTA Glass Tube

Tube

Target Data Initial 72 Hours % 72 Hours % Markers Points Cells/μΙ Cells/μΙ Difference Cells/μΙ Difference

CD3 12 111 1 11 16 0.5% 1107 0.4% CD4 12 766 755 1.4% 756 1.3%

[00149] Table 7. Percent difference of cell count recovery between day 7 glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for shipping and handling study of non-shipped tubes.

[00150] Table 8. Percent difference of cell count recovery between day 14 glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for shipping and handling study of non-shipped tubes.

[00151] Table 9. Percent difference of cell count recovery between day 7 glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for shipping and handling study of shipped tubes.

[00152] Table 10. Percent difference of cell count recovery between day 14 glass tube with immunophenotyping preservative reagent, polymeric coated tube with immunophenotyping preservative reagent and initial EDTA for shipping and handling study of shipped tubes.

[00153] Gas Permeability Testing

[00154] Gas permeability testing was performed on ten 9 ml and ten 4 ml blood collection tubes of the present teachings. The oxygen transmission rate (OTR) constant was measured to demonstrate the stability of the vacuum over 24 months closed-vial stability. The oxygen transmission rate constant of the blood collection tubes of the present teachings confirms the stability of the vacuum over 24 months product dating. FIG. 9(a) illustrates that the 9 ml blood collection tubes of the present teachings have an oxygen transmission rate 6 times lower than PET (polyethylene terephthalate) tubes. FIG. 9(b) illustrates that the 4 ml blood collection tubes of the present teachings have an oxygen transmission rate 5 times lower than PET (polyethylene terephthalate) tubes. FIG. 10(a) depicts the theoretical draw volume over product dating of the 9 ml blood collection tubes of the present teachings vs. PET tubes. There is a 6.3% loss compared to a 37.1 % loss in draw volume over 24 months. FIG. 10(b) depicts the theoretical draw volume over product dating of the 4 ml blood collection tubes of the present teachings vs. PET tubes. There is a 9.8% loss compared to a 50.1 % loss in draw volume over 24 months.

[00155] Moisture Transmission Testing

[00156] Moisture transmission testing was performed on PET tubes and blood collection tubes of the present teachings over 28 days at 18-25 °C, 30 ± 2 °C, and 50 ± 2 °C. These tests were performed to demonstrate that the blood collection tubes of the present teachings do not allow the stabilizing reagent to lose moisture over time. Moisture loss can cause the stabilizing reagent to become concentrated, which can produce excessive hemolysis. The 28-day moisture transmission study for all three temperature conditions demonstrates that the blood collection tubes of the present teachings do not allow the stabilizing reagent to lose significant moisture over time. Moisture loss (mg) over 28 days for the blood collection tubes of the present teachings vs. PET is displayed in FIG. 1 1 .

[00157] Results of a comparative analysis moisture vapor barrier study over 85 days at 22 °C, 30 °C and 50 °C of commercial plastic blood collection tubes and blood collection tubes of the present teachings are depicted in FIG. 12. The initial reagent fill volume is 200 μΙ_ for the blood collection tube of the present teachings, 300 μΙ_ for plastic blood collection tube # 1 and 1500 μΙ_ for plastic blood collection tube # 2. At a storage temperature of 22 °C, the blood collection tube of the present teachings had a reported loss of 1 .3 μΙ_ (« 0.65%), plastic blood collection tube # 1 had a reported loss of 41 .5 μΙ_ (« 13.8%) and plastic blood collection tube # 1 had a reported loss of 45.6 μΙ_ (« 3.0%). At a storage temperature of 30 °C, the blood collection tube of the present teachings had a reported loss of 7.1 μΙ_ (« 3.6%), plastic blood collection tube # 1 had a reported loss of 166.2 μΙ_ (~ 55.4%) and plastic blood collection tube # 2 had a reported loss of 207.5 μΙ_ (~ 13.8%). At a storage temperature of 50 °C, the blood collection tube of the present teachings had a reported loss of 31 .3 μΙ_ (~ 15.7%), plastic blood collection tube # 1 had a reported loss of 298 μΙ_ (« 99.3%) and plastic blood collection tube # 2 had a reported loss of 641 μΙ_ (~ 42.7%). The 85 day comparative analysis moisture vapor barrier study for all three temperature conditions demonstrates that the blood collection tubes of the present teachings do not allow the stabilizing reagent to lose significant moisture over time as compared to commercial plastic blood collection tubes.

[00158] Pressure Differential Testing

[00159] Pressure differential testing was performed on twelve blood collection tubes of the present teachings. This test satisfies FDA requirement 49 CFR §173.196(a)(6), which states that the primary receptacle or secondary packaging used for potentially infectious substances must be capable of withstanding, without leakage, an internal pressure producing a pressure differential of not less than 95 kPa. No physical issues were observed when the blood collection tubes of the present teachings were exposed to 95 kPa pressure differential for at least 30 minutes.

[00160] Twelve 9 ml blood collection tubes which are designed to draw in approximately 9 milliliters of a blood sample were subjected to a pressure differential test as described. A leak detection solution was added prior to testing. Each test sample was cleaned with a dry paper wipe to remove any leak detection solution. A dry paper wipe was secured around the closure of each test sample. Each test sample was placed into a hole in a corrugated board holder. Once all test samples had been placed into the corrugated board holder, the corrugated board holder was placed inside a vacuum chamber. The test samples were oriented with the closures down, ensuring the leak detection solution was in contact with the seal during the testing. The vacuum chamber was sealed, and the pressure inside the vacuum chamber was slowly reduced to at least 95 kPa below ambient air pressure. The test samples were exposed to the 95 kPa pressure differential for at least 30 minutes. The pressure inside the vacuum chamber was slowly increased to ambient air pressure, then the vacuum chamber was opened. The corrugated board holder was removed from the vacuum chamber, and each test sample was subjected to a visual inspection for leaks or any signs of damage (cracks, crazing, etc.).

[00161] A visual inspection was performed upon the completion of this test. No obvious physical issues (cracks, leaks, etc.) were observed upon the completion of this test. Thus, the sample collection tubes of the present teachings provide blood collect tubes which are compliant with established FDA standards. Therefore, the present teachings provide sample collection tubes in which collected samples may be shipped by air for testing and analysis.

[00162] International Safe Transit Association (ISTA) 1 A Testing

[00163] ISTA 1 A testing applies to packaged products weighing 150 lbs. or less and challenges the capability of the package and product to withstand transport hazards. ISTA 1A testing was performed on 12 twelve blood collection tubes of the present teachings. Acceptance criteria was a passing result for ISTA 1A (shock drop and fixed displacement vibration) testing. No physical issues were observed when the twelve blood collection tubes of the present teachings were subjected to ISTA 1 A (shock drop and fixed displacement vibration) testing.

[00164] Centrifuge Testing

[00165] Twenty blood-filled blood collection tubes of the present teachings were centrifuged at 3000 x g for 10 minutes in accordance with ISO 6710. Each tube was inspected for cracks or leakage after each centrifugation. Twenty tubes were sufficient to demonstrate the durability of the tubes under extreme centrifugation conditions. No cracks or leakage were detected after the filled blood collection tubes of the present teachings were centrifuged.

EXAMPLES

[00166] Example 1

[00167] In one example, the sample collection tube is a 4.0 ml blood collection tube.

The blood collection tube is about 13mm x 75mm. The fill tolerance volume is about

60.06 μΙ +/- 10% (54.58 μΙ to 66.70 μΙ/tube). The fill by weight is about 0.0708g +/- 10%

(0.0637g to 0.0779g/tube). The draw tolerance is about 4ml and may range from about

3.6 ml to about 4.4 ml. The vacuum expiration date is 24 months from the date of manufacture.

[00168] Example 2

[00169] In another example, the sample collection tube is a 10.0 ml blood collection tube for cell-free nucleic acid analysis. The blood collection tube is about 16mm x 100 mm. The fill tolerance volume is about 200.0 μΙ +/- 10% (180 μΙ to 220 μΙ/tube). The fill by weight is about 0.249g +/- 10% (0.2241 g to 0.2739g/tube). The draw tolerance is about 10 ml and may range from about 9.0 ml to about 1 1 .0 ml. The vacuum expiration date is 24 months from the date of manufacture.

[00170] Example 3

[00171] An exemplary sample collection tube may be a 9.0 ml blood collection tube. The blood collection tube is about 16mm X 100 mm. The fill tolerance volume is about 180.0 μΙ +/- 10% (162 μΙ to about 198 μΙ). The fill by weight is about 0.2241 g +/- 10% (0.2017g to 0.2564g/tube). The draw tolerance is about 9 ml and may range from +/- 10% (e.g. 8.1 ml to 9.9 ml). The draw height may be about 80mm to about 84 mm. The vacuum expiration date is 24 months from the date of manufacture.

[00172] As used herein, unless otherwise stated, the teachings envision that any member of a genus (list) may be excluded from the genus; and/or any member of a Markush grouping may be excluded from the grouping.

[00173] Unless otherwise stated, any numerical values recited herein include all values from the lower value to the upper value in increments of one unit provided that there is a separation of at least 2 units between any lower value and any higher value. As an example, if it is stated that the amount of a component, a property, or a value of a process variable such as, for example, temperature, pressure, time and the like is, for example, from 1 to 90, preferably from 20 to 80, more preferably from 30 to 70, it is intended that intermediate range values such as (for example, 15 to 85, 22 to 68, 43 to 51 , 30 to 32 etc.) are within the teachings of this specification. Likewise, individual intermediate values are also within the present teachings. For values which are less than one, one unit is considered to be 0.0001 , 0.001 , 0.01 or 0.1 as appropriate. These are only examples of what is specifically intended and all possible combinations of numerical values between the lowest value and the highest value enumerated are to be considered to be expressly stated in this application in a similar manner. As can be seen, the comparative teaching of amounts expressed as weight/volume percent for two or more ingredients also encompasses relative weight proportions of the two or more ingredients to each other, even if not expressly stated. For example, if a teaching recites 2% A, and 5% B, then the teaching also encompasses a weight ratio of A: B of 2:5. Unless otherwise stated, all ranges include both endpoints and all numbers between the endpoints. The use of "about" or "approximately" in connection with a range applies to both ends of the range. Thus, "about 20 to 30" is intended to cover "about 20 to about 30", inclusive of at least the specified endpoints.

[00174] The disclosures of all articles and references, including patent applications and publications, are incorporated by reference for all purposes.

[00175] The term "consisting essentially of to describe a combination shall include the elements, ingredients, components or steps identified, and such other elements ingredients, components or steps that do not materially affect the basic and novel characteristics of the combination. The use of the terms "comprising" or "including" to describe combinations of elements, ingredients, components or steps herein also contemplates embodiments that consist essentially of (namely, the presence of any additional elements, ingredients, components or steps, does not materially affect the properties and/or benefits derived from the teachings; or even consist of the elements, ingredients, components or steps.

[00176] Plural elements, ingredients, components or steps can be provided by a single integrated element, ingredient, component or step. Alternatively, a single integrated element, ingredient, component or step might be divided into separate plural elements, ingredients, components or steps. The disclosure of "a" or "one" to describe an element, ingredient, component or step is not intended to foreclose additional elements, ingredients, components or steps. All references herein to elements or metals belonging to a certain Group refer to the Periodic Table of the Elements published and copyrighted by CRC Press, Inc., 1989. Any reference to the Group or Groups shall be to the Group or Groups as reflected in this Periodic Table of the Elements using the lUPAC system for numbering groups.

[00177] It is understood that the above description is intended to be illustrative and not restrictive. Many embodiments as well as many applications besides the examples provided will be apparent to those of skill in the art upon reading the above description. The scope of the teachings should, therefore, be determined not with reference to the above description, but should instead be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled. The disclosures of all articles and references, including patent applications and publications, are incorporated by reference for all purposes. The omission in the following claims of any aspect of subject matter that is disclosed herein is not a disclaimer of such subject matter, nor should it be regarded that the inventors did not consider such subject matter to be part of the disclosed inventive subject matter.

[00178] The explanations and illustrations presented herein are intended to acquaint others skilled in the art with the teachings, its principles, and its practical application. Those skilled in the art may adapt and apply the teachings in its numerous forms, as may be best suited to the requirements of a particular use. Accordingly, the specific embodiments of the present teachings as set forth are not intended as being exhaustive or limiting of the teachings. The scope of the teachings should, therefore, be determined not with reference to the above description, but should instead be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled. The disclosures of all articles and references, including patent applications and publications, are incorporated by reference for all purposes. Other combinations are also possible as will be gleaned from the following claims, which are also hereby incorporated by reference into this written description.

Claims

CLAIMS What is claimed is:

1 . A blood collection sample tube comprising: a tube having an enclosed base, a coextensive elongated side wall extending from the base and terminating at an open end, and defining a hollow chamber having an inner wall, the hollow chamber being configured for collecting a blood sample, at least the elongated side wall of the tube being made of a material including a thermoplastic polymeric material having a high moisture barrier and low moisture absorption rate, optical transparency to enable viewing a sample within the tube and chemical resistance; and optionally a transparent silicon-containing coating on at a majority of the side wall of the tube; and an elastomeric stopper; wherein prior to collecting the blood sample, the hollow chamber of the tube is in an evacuated condition relative to an ambient pressure, and the hollow chamber is partially filled with a reagent in an initial state selected from a solid, a liquid or a gel, the reagent including an optional anticoagulant, and a preservative composition adapted for stabilizing blood cells of the blood sample for enabling isolation of a nucleic acid or a rare cell circulating in the blood sample and/or flow cytometry testing of the blood sample; and wherein the reagent is capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C.

2. The blood collection tube of claim 1 , wherein the polymeric material includes a cyclic polyolefin.

3. The blood collection tube of claim 1 or claim 2, wherein the polymeric material includes cyclic polyolefin copolymer (COC).

4. The blood collection tube of any of the preceding claims, wherein the tube is part of a kit adapted for a non-invasive testing of a maternal blood sample of a pregnant woman.

5. The blood collection tube of any of the preceding claims, wherein the stopper includes bromobutyl rubber.

6. The blood collection tube of any of the preceding claims, wherein the stopper of the tube includes a silicone oil coating over at least a portion of its outer surface that contacts the inner wall of the tube.

7. The blood collection tube of any of the preceding claims, wherein the tube has an outer diameter, as measured at the coextensive elongated side wall adjacent the open end, to length (D x L) dimension of about 13 mm x 75 mm.

8. The blood collection tube of any of the preceding claims, wherein the tube has an outer diameter, as measured at the coextensive elongated side wall adjacent the open end, to length (D x L) dimension of about 16 mm X 100 mm.

9. The blood collection tube of any of the preceding claims, wherein the tube includes a reagent fill tolerance volume of about 54 μΙ to about 66 μΙ.

10. The blood collection tube of any of the preceding claims, wherein the tube includes a reagent fill tolerance volume of about 60 μΙ.

1 1 . The blood collection tube of any of the preceding claims, wherein the tube includes a reagent fill tolerance volume of about 160 μΙ to about 220 μΙ.

12. The blood collection tube of any of the preceding claims, wherein the tube includes a reagent fill tolerance volume of about 180 μΙ.

13. The blood collection tube of any of the preceding claims, wherein the tube includes a reagent fill by weight of plus or minus 10% of 0.0700 g.

14. The blood collection tube of any of the preceding claims, wherein the tube includes a reagent fill by weight of plus or minus 10% of 0.250 g.

15. The blood collection tube of any of the preceding claims, wherein the tube includes a draw tolerance of about 3 ml to about 5 ml.

16. The blood collection tube of any of the preceding claims, wherein the tube includes a draw tolerance of about 4 ml.

17. The blood collection tube of any of the preceding claims, wherein the tube includes a draw tolerance of about 7 ml to about 13 ml.

18. The blood collection tube of any of the preceding claims, wherein the tube includes a draw tolerance of about 10 ml.

19. The blood collection tube of claim of any of the preceding claims, wherein the preservative composition, contains less than about 20 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition.

20. The blood collection tube of any of the preceding claims, wherein the preservative composition contains less than about 15 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition.

21 . The blood collection tube of any of the preceding claims, wherein the preservative composition contains less than about 10 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition.

22. The blood collection tube of any of the preceding claims, wherein the preservative composition contains less than about 5 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition.

23. The blood collection tube of any of the preceding claims, wherein the preservative composition contains at least about 100 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition.

24. The blood collection tube of any of the preceding claims, wherein the preservative composition contains at least about 1000 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition.

25. The blood collection tube of any of the preceding claims, wherein the preservative composition contains at least about 10,000 parts per million (ppm) of an aldehyde per 100 μΙ of the preservative composition.

26. The blood collection tube of any of the preceding claims, wherein the aldehyde includes formaldehyde.

27. The blood collection tube of any of the preceding claims, wherein the polymeric material has a moisture vapor transmission rate of about 0.023 g ^■ mm/ m^{2 ■} d to about 0.045 g ^■ mm/ m^{2 ■} d at 23 °C and 85% relative humidity, as measured by DIN 53 122.

28. The blood collection tube of any of the preceding claims, wherein the polymeric material has a tensile strength of about 60 MPa to about 63 MPa, as measured by ISO 527, parts 1 and 2.

29. The blood collection tube of any of the preceding claims, wherein the polymeric material has a tensile modulus of about 2300 MPa to about 2900 MPa, as measured by ISO 527, parts 1 and 2.

30. The blood collection tube of any of the preceding claims, wherein the polymeric material has an impact strength (Charpy Impact Unnotched) of about 20 kJ/m2 as measured by ISO 179/1 ell.

31 . The blood collection tube of any of the preceding claims, wherein the polymeric material has a light transmission of at least about 90%.

32. The blood collection tube of any of the preceding claims, wherein the polymeric material has a mold shrinkage of about 0.1 % to about 0.7%.

33. The blood collection tube of any of the preceding claims, wherein the polymeric material has a birefringence of less than 20 nm according to ASTM D4093-95.

34. The blood collection tube of any of the preceding claims, wherein the polymeric material has a glass transition temperature of about 70 °C to about 140 °C as measured by differential scanning calorimetry (DSC).

35. The blood collection tube of any of the preceding claims, wherein the polymeric material and coating each have coefficients of thermal expansion that are within about twenty percent of each other.

36. The blood collection tube of any of the preceding claims, wherein the polymeric material includes a homopolymer or copolymer that includes polyethylene,

polypropylene or both.

37. The blood collection tube of any of the preceding claims, wherein the polymeric material includes a cyclic moiety.

38. The blood collection tube of any of the preceding claims, wherein the polymeric material includes a polyester.

39. The blood collection tube of any of the preceding claims, wherein the preservative is selected from the group consisting of formaldehyde, diazolidinyl urea (DU), imidazolidinyl urea (IDU), dimethylol urea, 2-bromo-2-nitropropane-1 ,3-diol, 5- hydroxymethoxymethyl-1 -aza-3,7-dioxabicyclo (3.3.0)octane and 5-hydroxymethyl-1 - aza-3,7-dioxabicyclo (3.3.0)octane and 5-hydroxypoly [methyleneoxy]methyl-1 -aza-3,7- dioxabicyclo (3.3.0)octane, bicyclic oxazolidines, DMDM hydantoin, sodium

hydroxymethylglycinate, hexamethylenetetramine chloroallyl chloride, biocides, a water- soluble zinc salt and any combination thereof.

40. The blood collection tube of any of the preceding claims, wherein the anticoagulant is EDTA, anticoagulant citrate dextrose-A, anticoagulant citrate dextrose-B, heparin, or combinations thereof.

41 . The blood collection tube of any of the preceding claims, wherein the tube is suitable for storing a blood sample for a period of at least about 14 days.

42. The blood collection tube of any of the preceding claims, including an adhesive- backed label adhered to an outer wall of the tube, wherein the adhesive is an acrylic based adhesive.

43. The blood collection tube of any of the preceding claims, wherein the reagent in the tube includes about 15% to about 70% by volume of the preservative composition prior to blood draw.

44. The blood collection tube of any of the preceding claims, wherein the reagent in the tube includes about 15% to about 70% by volume of imidazolidinyl urea (IDU) prior to blood draw.

45. The blood collection tube of any of the preceding claims, wherein the reagent in the tube includes about 5% to about 20% by volume of anticoagulant prior to blood draw.

46. The blood collection tube of any of the preceding claims, wherein the reagent in the tube includes about 1 % to about 20% of polysaccharide prior to blood draw.

47. The blood collection tube of any of the preceding claims, wherein reagent in the tube includes about 1 % to about 10% of an agent in addition to the preservative and an anticoagulant prior to blood draw.

48. The blood collection tube of any of the preceding claims, wherein the tube includes an amount of aldehyde reaction agent for reacting with free formaldehyde present in the reagent sufficient to result in an aldehyde amount below 15,000 ppm per 100 μΙ of reagent prior to blood draw.

49. The blood collection tube of any of the preceding claims, wherein the tube includes an amount of aldehyde reaction agent containing a primary amine for reacting with free formaldehyde present in the reagent to result in an aldehyde amount below 100 ppm per 100 μΙ of reagent prior to blood draw.

50. The blood collection tube of any of the preceding claims, wherein the tube includes an amount of aldehyde reaction agent, which aldehyde reaction agent is selected from one or any combination of tris, glycine, or a derivative of either or both, for reacting with free formaldehyde present in the reagent to result in an aldehyde amount below 20 ppm per 100 μΙ of reagent prior to blood draw.

51 . The blood collection tube of any of the preceding claims, wherein the aldehyde reaction agent is present in a ratio (by weight) relative to the preservative composition of about 1 :20 to about 1 :1 .

52. The blood collection tube of any of the preceding claims, wherein the aldehyde reaction agent is present in a ratio (by weight) relative to the anticoagulant of about 1 :25 to about 5:1 .

53. The blood collection tube of any of the preceding claims, wherein the preservative composition is present in a ratio (by weight) relative to the anticoagulant of about 1 : 10 to about 15: 1.

54. The blood collection tube of any of the preceding claims, wherein the reagent is present in the tube as a liquid in an amount less than about 10% by volume of the tube but greater than about 0.1 % by volume.

55. The blood collection tube of any of the preceding claims, wherein the reagent is present in the tube as a liquid in an amount less than about 5% by volume of the tube but greater than about 0.1 % by volume of the tube.

56. The blood collection tube of any of the preceding claims, wherein the reagent is present in the tube in an amount less than about 3% by volume of the tube but greater than about 0.1 % by volume of the tube.

57. The blood collection tube of any of the preceding claims, wherein the tube includes a reagent with one or more ingredients and wherein a ratio of the one or more ingredients is about 5: 1 to about 15: 1 .

58. The blood collection tube of any of the preceding claims, wherein the tube includes imidazolidinyl urea (IDU) and glycine and a ratio of imidazolidinyl urea (IDU) to glycine is about 10: 1.

59. The blood collection tube of any of the preceding claims, wherein the reagent is capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C when the tube is subjected to relative humidity of up to about least about 50%.

60. The blood collection tube of any of the preceding claims, wherein the reagent is capable of retaining its initial state for a period of at least one month over a temperature range of about 2 °C to about 30 °C when the tube is subjected to relative humidity of up to about 75%.

61 . The blood collection tube of any of the preceding claims, wherein the reagent in the tube includes about 60% to about 90% by volume diazolidinyl urea (DU) prior to blood draw.

62. The blood collection tube of any of the preceding claims, wherein the reagent in the tube includes about 15% to about 35% of EDTA prior to blood draw.