CN216386455U - Collection system of dry blood spot is gathered to ration - Google Patents

Abstract

The utility model discloses a collection device for quantitatively collecting dried blood spots, which comprises a sample collection ring with two through ends, a sample carrying piece and a quantitative capillary tube for sampling, wherein the sample carrying piece is detachably arranged in the sample collection ring. The utility model can quantitatively collect blood samples, can directly seal and send the samples without drying in the air, avoids punching and sampling, reduces cross contamination, improves the accuracy of the quantitative detection result of dried blood spots, and integrally completes the operations of quantitative collection and storage of the samples, input of sample information and sending of the samples through the device.

Description

Collection system of dry blood spot is gathered to ration

Technical Field

The utility model relates to the technical field of dry blood spot detection, in particular to a collecting device for quantitatively collecting dry blood spots.

Background

Dry Blood Spot (DBS) detection is a method in which a clinical biological sample is dropped onto a filter paper sheet, Dried and stored, and then subjected to solvent extraction to detect analytes in the dry Blood Spot, the technology uses the filter paper as a carrier, and is widely used for qualitative detection of nucleic acid genetic materials, qualitative and quantitative detection of biomarkers, monitoring of drug concentration and the like, with the development of sequencing technology and mass spectrometry technology, the application of the dry Blood Spot in clinical disease detection and screening projects is more and more extensive, but the current dry Blood Spot collection card needs to be Dried in the shade for two to three hours in the Blood Spot preparation process, so that not only is the sample preparation efficiency affected, but also cross contamination is easily generated in the process; manual punching is needed in the use process of the dry blood spots, so that time and labor are consumed, and cross contamination and confusion among samples are easy to generate; in addition, due to blood sampling volume effect, hematocrit effect (HCT) and chromatography effect, the components at the center and the edge of the same blood spot are not uniform, holes are punched at different parts of the blood spot, and the content of the components is not uniform, so that the detection error is large and the reproducibility is poor.

Patent specification No. CN111474257A discloses a quantitative collection device for dried blood spots, comprising: the device comprises a sample collecting pipe, sample collecting filter paper and a quantitative blood collecting pipe; the sample collection tube comprises a tube body and a tube cover; the pipe body is a straight pipe, and the pipe cover is used for sealing the pipe orifice; the quantitative blood collection tube comprises a quantitative capillary tube and a fixing piece; the quantitative capillary tube is fixed on the fixing piece and provided with a blood sampling end which is used for sampling blood and is exposed out of the fixing piece; the fixing piece is clamped on the pipe orifice, and the quantitative capillary tube extends into the pipe body; the sample collection filter paper is arranged in the tube body, and the blood sampling end is abutted against the sample collection filter paper.

The patent specification with the publication number of CN206192677U discloses a collection tube type dried blood spot sampling device, which comprises a collection tube main body with two through ends, wherein the top of the collection tube main body is provided with a top sealing film which can be pressed and broken, an information identification code is printed on the top sealing film, and the bottom of the collection tube main body is provided with matched filter paper.

Above-mentioned two kinds of schemes all have the blood spot to dry the problem that the time is longer, influence system appearance efficiency.

SUMMERY OF THE UTILITY MODEL

The utility model aims to provide a collection device for quantitatively collecting dried blood spots, which can quantitatively collect blood samples, can directly seal and send the samples without drying in the air, avoids punching and sampling, reduces cross contamination, improves the accuracy of the quantitative detection result of the dried blood spots, and integrally completes the operations of quantitative collection, storage, sample information input and sample sending of the samples through the device.

The utility model provides a collection system of quantitative collection dry blood spot, includes the penetrating sample collection ring in both ends, carries the appearance piece and is used for the ration capillary of sampling, carry appearance piece detachably to set up in the sample sampling intra-annular.

In this scheme, carry out quantitative sampling with the ration capillary, the blood sample of ration collection is whole to be shifted to on carrying the sample piece, and the whole sample piece carries the sample piece and puts into the extraction tube and extract the analyte, can avoid the sample to carry the sample on carrying the sample piece the quantitative result that the uneven distribution arouses undulant big, the problem that the degree of accuracy is low.

Preferably, the sample loading piece comprises a plurality of pieces arranged in the sample sampling ring, and the pieces are arranged at intervals along the axial direction of the sample sampling ring.

Preferably, a fixing clamp for fixing the sample carrying sheet is arranged on the inner wall of the sample collecting ring; the fixation clamp comprises a plurality of sample collection ring inner walls which are uniformly distributed along the circumferential direction, at least one of the sample collection ring inner walls is telescopic, and a hard sample carrying piece is conveniently placed into the sample collection ring.

Further preferably, the end part of the fixing clip for fixing is provided with a bayonet for inserting the sample carrying piece.

Preferably, a plurality of uniformly distributed hollows are arranged along the circumferential direction of the ring wall of the sample collecting ring. Three hollows are arranged in the middle of the sample collecting ring, so that the blood spots on the sample carrying sheet can be conveniently ventilated and dried.

Preferably, the sample carrier is all-cotton fiber filter paper, glass fiber filter paper or diatomite. The waterman 903 filter paper can be adopted specifically, and the sample carrying sheet can also be a wafer pressed by other appropriate materials, such as a wafer pressed by inorganic powder; the diameter of the sample loading piece is 3mm to 12mm, and preferably 3mm to 8 mm.

Preferably, the sampling device further comprises a bracket, wherein a plurality of concave cavities for placing a plurality of sample sampling rings are arranged on the bracket. The sample sampling rings are marked with numbers, and the side surface of the bracket is provided with an area capable of being stuck with a sample information bar code for sticking the sample information bar code. The material of the sample collection ring and the material of the bracket are the same plastic material, and the plastic material is preferably transparent.

Preferably, the support is further provided with a placing cavity for placing the quantitative capillary, one end of the placing cavity is closed, the other end of the placing cavity is opened, and the opening end of the placing cavity is provided with an end cover for closing. The placing cavity is horizontally placed at the front end of the bracket, and the length of the placing cavity is slightly longer than that of the quantitative capillary.

Preferably, the scaffold further comprises a sealing bag for placing the scaffold, and a drying agent is arranged in the sealing bag. The sealing bag mouth is provided with a concave-convex sealing clamping strip which can be opened and sealed. The sealing bag can be made of transparent PE material, and if the measured sample is sensitive to light, the sealing bag can be light-shading.

A collection method adopts the collection device to collect dry blood spots, and comprises the following steps:

s1: the blood sampling end of the quantitative capillary is close to and contacted with blood, and the blood is filled in the whole quantitative capillary or reaches corresponding volume scales by the capillary force;

s2: after a blood sample is collected, the blood collecting end of the quantitative capillary is contacted with the sample carrying sheet, and the blood sample transfers all the blood in the quantitative capillary to the sample carrying sheet in the sample collecting ring by means of air pressure and the suction force of the sample carrying sheet;

s3: and (4) sealing and storing the sample collecting ring.

Before packaging, the bar code of the blood sample information is pasted on the bracket, and the quantitative collection device of the dry blood spots is placed in a proper environment for storage or transportation after being sealed. For some samples sensitive to air oxidation, the samples can be sealed after being filled with nitrogen before sealing, so that the stability of the samples is improved.

Preferably, when analyzing the sample, one or more of the sample-carrying sheets is/are taken out and put into the sample extraction tube, the extraction liquid is directly added to extract the analyte, and the extracted analyte is sent for detection.

The utility model has the beneficial effects that:

(1) the quantitative capillary is adopted for quantitative sampling, all blood samples which are quantitatively collected are transferred to the sample carrying piece, the whole sample carrying piece is placed into the extraction tube for extracting the analyte, and the problems of large fluctuation of quantitative results and low accuracy caused by uneven distribution of the samples on the sample carrying piece can be avoided.

(2) The quantitative capillary is adopted to collect blood samples, so that the samples can be quantitatively transferred onto the sample loading piece, and the detection result is more accurate.

(3) The blood sample need not dry after adding on carrying the sample piece, can direct sealed save or transport, has reduced the latency who dries the process, has improved the effect, has also reduced the sample pollution's of the process of drying possibility simultaneously.

(4) Before the samples are extracted, each sample is not required to be punched, so that the working intensity is reduced, the efficiency is improved, and the cross contamination caused by punching is reduced.

(5) For samples with different polarities, sample carrying sheets made of different materials can be selected, so that the extraction recovery degree can be improved, and the detection sensitivity can be improved.

Drawings

FIG. 1 is a schematic structural view of a stent according to the present invention;

FIG. 2 is a schematic diagram of a sample acquisition loop;

FIG. 3 is a schematic view of the structure of the portion A in FIG. 2;

FIG. 4 is a graph showing the drying times for different sampling amounts in examples 1 to 3;

FIG. 5 is a schematic view showing the relationship between the concentration of the analyte and the amount of sample in example 4;

FIG. 6 is a schematic diagram showing the relationship between the number of different loading pieces and the concentration of the analyte in example 6;

FIG. 7 is a graph showing the stability of the sample of example 7 over 12 days;

FIG. 8 is a typical spectrum of mass spectrometric detection of 25-hydroxyvitamin D.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

As shown in fig. 1-3, a collection device for quantitatively collecting dry blood spots comprises a sealing bag, a support 1, a sample collection ring 2, a sample carrying sheet 3 and a quantitative capillary tube, wherein the support 1 is used for accommodating the sample collection ring 2, the sample carrying sheet 3 and the quantitative capillary tube, and is packaged in the sealing bag for sealing after sampling.

Wherein, the sample sampling ring 2 is cylindric and both ends are penetrating, is provided with a plurality of sample carriers 3 of interval arrangement along its axial in the sample sampling ring 2, and carries sample carrier 3 and sample sampling ring 2's port parallel arrangement.

In this embodiment, two sample loading pieces 3 are provided, which are respectively close to one quarter of the two ports of the sample sampling ring 2, and two groups of fixing clamps 21 are provided on the inner wall of the sample sampling ring 2 for fixing the sample loading pieces 3; in this embodiment, three fixing clips 21 are specifically arranged in each group, and are uniformly distributed along the circumferential direction of the inner wall of the sample collection ring 2, and one of the fixing clips is retractable, the retractable principle is conventional, or the fixing clip 21 is directly made of an elastic material, a bayonet 211 is arranged at one end of the fixing clip 21 for fixing, and the diameter of the sample carrying sheet 3 is smaller than the inner diameter of the sample collection ring 2, so that the sample carrying sheet is convenient to put in; when the fixing clamp is placed, the telescopic fixing clamp 21 is pressed, the edge of the sample loading piece 3 is placed in the bayonet 211 of the fixing clamp 21, and the telescopic fixing clamp 21 is reset to complete fixing.

In this embodiment, three evenly distributed hollows 22 are circumferentially arranged in the middle of the annular wall of the sample collection ring 2, so that the blood spots on the sample carrying sheet 3 can be conveniently ventilated and dried. The sample carrying sheet 3 can be made of all-cotton fiber filter paper, glass fiber filter paper or diatomite, or can be made of other discs made of other appropriate materials by pressing, and the diameter is 3-8 mm.

In this embodiment, the bracket 1 is provided with cavities 11 distributed in 2 × 3, each cavity 11 is provided with a corresponding sample collection ring 2, the sample collection rings 2 are marked with numbers, and the side surface of the bracket 1 is provided with an area capable of being pasted with a sample information bar code for pasting the sample information bar code.

The front end of the bracket 1 is provided with a horizontally placed placing cavity 12, the length of which is slightly longer than that of the quantitative capillary, and the opening end is provided with an end cover 13 for closing.

In the embodiment, the drying agent is arranged in the sealing bag, and the opening of the sealing bag is provided with the concave-convex sealing clamping strip which can be opened and sealed. The sealing bag can be made of transparent PE material, and if the measured sample is sensitive to light, the sealing bag can be light-shading.

A collection method for collecting dry blood spots by adopting the collection device comprises the following steps:

s1: opening the sealing bag, taking out the quantitative capillary tube and the sample sampling ring with the sample carrying sheet from the bracket, enabling the blood sampling end of the quantitative capillary tube to be close to and contact with blood, and filling the whole quantitative capillary tube or corresponding volume scales with the blood by capillary force;

s2: after a blood sample is collected, the blood collecting end of the quantitative capillary is contacted with the circle center of the sample carrying sheet, and the blood sample transfers all blood samples in the quantitative capillary to the sample carrying sheet by means of air pressure and the suction force of the sample carrying sheet;

s3: sticking a blood sample information bar code on the bracket;

s4: after the quantitative capillary tube and the sample sampling ring are completely stored, the whole body is placed into a sealing bag filled with a drying agent, air in the bag is squeezed out, the bag is sealed, and the dry blood spot collecting device is placed in a proper environment for storage or transportation;

s5: when the sample is analyzed, the sealing bag is opened, the bracket is taken out, after the sample information is input by scanning codes, one or more sample carrying pieces are taken out and put into the extraction tube, the extraction liquid is added to extract the analyte, and the extracted analyte is sent for inspection.

Examples 1 to 3

The quantitative collection device is used for quantitatively collecting 20, 30 and 40 mu L of whole blood respectively, and the quality of the dried blood spots is measured after the whole blood is placed at room temperature for 2h, 4h, 6h and 24h after collection is finished, and the result is shown in figure 4, wherein the quality is the total weight of the dried blood spots and the filter paper sheet.

It can be seen from figure 4 that the dried blood spot weight did not change substantially after 6 hours, indicating that the collection device had a drying time of 6 hours and more at room temperature.

Example 4

The quantitative collection device is used for quantitatively collecting 10, 20, 30 and 40 mu L of whole blood respectively, and after collection is finished, the whole blood is placed at room temperature overnight, and then the concentration of 25-hydroxyvitamin D in dry blood spots with different blood collection amounts is measured.

The detection conditions were as follows:

1. sample processing

Taking out the dried blood spot sample and placing the dried blood spot sample in a 96-well plate;

add 100. mu.L of reagent A to each well separately;

then 200. mu.L of reagent B was added to each well;

then 40uL of VD-IS with the concentration of 10ng/mL IS added into the solution, and the solution IS continuously shaken for more than 30 min;

transferring the solution in each well to a 96-well SLE plate, and standing for about 5 min;

adding 400uL of reagent C into each hole, and standing for about 5 min;

adding 400uL of reagent C into each hole, and standing for about 5 min;

drying by nitrogen;

adding 60 mu L of reagent D into each hole with the sample, and vortex mixing for 2 min;

derivatizing at 50 deg.C for 30 min;

adding 40 mu L of reagent B into each hole, and mixing uniformly for 2min by vortex;

drying by nitrogen;

adding 100 mu L of reagent E into each hole with the sample, and mixing uniformly for 2min by vortex;

and (3) placing the 96-hole plate with the silica gel cover plate into an automatic sample injector, and starting a program to perform sample injection detection.

2. Liquid phase conditions

A chromatographic column: BEH C18, 3.0 × 50mm (waters);

mobile phase A: an aqueous solution containing 0.1% formic acid;

mobile phase B: acetonitrile solution containing 0.1% formic acid;

the sample injection amount is 5 mu L;

flow rate: 0.4 mL/min;

column temperature: 45 ℃;

sample injector temperature: 15 ℃;

elution gradient:

3. conditions of Mass Spectrometry

Source parameter

MRM ion channels

Test substance/internal standard	MRM	Taper hole voltage (V)	Collision energy (eV)
				VD2	570.4/298.2	10	8
VD2-IS	573.4/301.4	10	8
				VD3	558.4/298.2	4	8
VD3-IS	564.3/298.4	4	8

Under different sample loading conditions, the detection result of 25-hydroxyvitamin D in blood spots is shown in the following table 1, and a typical spectrum of 25-hydroxyvitamin D detected by liquid chromatography-tandem mass spectrometry is shown in fig. 8.

TABLE 1 results of 25-hydroxyvitamin D assay in dry blood spots of different loading amounts

Unit: ng/mL

As can be seen from the above table, the concentrations are substantially proportional to the sample amounts at the sample amounts of 10, 20 and 30. mu.L, and the concentrations are much smaller than the theoretical concentrations at the sample amount of 40. mu.L. The above-mentioned detection concentrations of 20, 30, and 40. mu.L in the sample amount were divided by the detection concentration of 10. mu.L in the sample amount, and this ratio was plotted as the y-axis and the theoretical ratio as the x-axis to obtain FIG. 5.

It can be seen from fig. 5 that the actual ratio of the 40 μ L sample size is much lower than the theoretical ratio (the inflection point appears at the fourth point in the figure). In summary, the sample loading capacity of the single loading device of the sampling device described herein preferably does not exceed 30 μ L.

Example 5

In this example, the concentration of the analyte in the whole blood sample (i.e., the whole blood from which the dried blood spot was prepared) was measured simultaneously, and the difference in the concentrations of the analyte in the two samples was compared, as shown in Table 2 below.

TABLE 2 recovery of dried blood spot samples

From table 2, it can be seen that the recovery rates are all greater than 100%, which indicates that the analyte in the dried blood spot can be completely extracted, and indicates that the sampling device of the present invention does not produce a large adsorption to the analyte.

Example 6

The quantitative collecting device can be provided with different sample carrying numbers (one, two, three and four), each sample is respectively used for quantitatively collecting 20 mu L of whole blood, and after the collection is finished, the whole blood is placed overnight at room temperature, and then the concentration of 25-hydroxyvitamin D in dry blood spots with different sample carrying numbers is measured. The results of the measurements are shown in Table 3 below.

TABLE 3 results of 25-hydroxyvitamin D assay for different numbers of sample-carrying tablets

Fig. 6 shows a scatter diagram obtained by dividing the detected concentrations at 1, 2, 3, and 4 sample carriers in the table by the detected concentration at 1 sample carrier, and plotting the ratio as the y-axis and the theoretical ratio as the x-axis.

As can be seen from Table 3 and FIG. 6, the number of the specimen slides is proportional to the concentration of the analyte, indicating that the sampling device can increase the sampling amount by increasing the number of the specimen slides.

Example 7

After 20 mu L of the sample is sampled by using the quantitative collecting device, the sample is respectively placed for 1 hour, 6 hours, 1 day, 4 days, 7 days and 12 days at room temperature and then uniformly treated, and whether the 25-hydroxy vitamin D in the dried blood spots is stable or not is measured. The stability of the samples is shown in figure 7.

As can be seen from fig. 7, the stability of the sample is good within 12 days, and the sample collected by the collecting device is considered to be capable of being stored at room temperature for 12 days.

Example 8

In this example, a sample carrier sheet made of diatomaceous earth was used in place of the original filter paper sheet, and the recovery of the sample using diatomaceous earth was as shown in Table 4 below.

TABLE 4 recovery of dried blood spot samples

From table 2, it can be seen that the recovery rate is between 89% and 104%, which indicates that the analyte in the dried blood spot of the sample-carrying sheet prepared by using diatomite can be completely extracted, and indicates that the sampling device of the present invention can also use diatomite as the sample-carrying sheet.

Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes in the embodiments and/or modifications of the utility model can be made, and equivalents and modifications of some features of the utility model can be made without departing from the spirit and scope of the utility model.

Claims (6)

1. The collecting device for quantitatively collecting the dried blood spots is characterized by comprising a sample collecting ring with two through ends, a sample carrying sheet and a quantitative capillary tube for sampling, wherein the sample carrying sheet is detachably arranged in the sample collecting ring; the inner wall of the sample collecting ring is provided with a fixing clamp for fixing the sample carrying piece; the fixing clamps comprise a plurality of fixing clamps which are uniformly distributed along the circumferential direction of the inner wall of the sample collecting ring, and at least one of the fixing clamps is telescopic; and a plurality of uniformly distributed hollows are arranged along the circumferential direction of the ring wall of the sample collecting ring.

2. The collection device of claim 1, wherein the sample-carrying sheet comprises a plurality of sheets disposed within the sample sampling ring and spaced axially along the sample sampling ring.

3. The collecting device as claimed in claim 1, wherein the end of the fixing clip for fixing is provided with a bayonet for inserting the sample carrier.

4. The collecting device as claimed in any one of claims 1 to 3, further comprising a rack, wherein the rack is provided with a plurality of cavities for placing a plurality of sample sampling rings.

5. The collection device according to claim 4, wherein the holder is further provided with a placing cavity for placing the quantitative capillary, one end of the placing cavity is closed, the other end of the placing cavity is open, and the open end is provided with an end cover for closing.

6. The collection device of claim 4, further comprising a sealed bag for holding the scaffold, wherein a desiccant is disposed within the sealed bag.

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