RU2616242C1 - Device for collecting, storage and transportation of biological material of living organisms and plants - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: device for the collection, storage and transport of biological materials and living organisms of plants comprising a carrier card with a sorption material placed in it, adapted to be applied to him the biological material containing the sample components, and its subsequent drying. The carrier card includes a card device made of coated paper weighing 300 g/m² and stiffness (MD) to 15 mNm rectangular form with two bends vertically and with the possibility of adding it in three layers, with the lower inner layer formed after folding the rectangle at two vertical folds, the membrane is rigidly fixed sorption. Sorption absorbent membrane is made of porous absorbing material is not less than 150 l of fluid from the fluid uptake rate of at least 1 cm/s and the capillary rise of at least 20 mm/min.

EFFECT: simplification of the design of the device, while expanding its scope, as well as increasing the sensitivity of laboratory tests.

5 cl, 2 dwg, 2 tbl, 6 ex

Description

The invention relates to the field of medicine, veterinary medicine, agriculture and can be used for the collection, storage and transportation of biological materials of living organisms and plants.

Currently, the use of sorbents for the collection, storage and transportation of biological materials is expanding rapidly, especially in medical, veterinary and agricultural practice.

The collection and transportation of biological material is the initial and one of the most critical stages for subsequent laboratory studies, since violation of the rules for the collection of biological material, obtaining unrepresentative clinical samples, and incorrect and untimely delivery affect the results of the study.

The essence of the collection, storage and transportation of biological materials is the collection of a certain amount of material, since its excess or deficiency affects the quality of DNA extraction and contribute to its degradation during storage for transportation. In addition, the sterility of both the collection of biological material and its storage and transportation must be respected.

A known container for a blood sample containing a container with a sealing lid, while in order to simplify the quantitative taking of a blood sample and increase the shelf life, a lid for attaching absorbent paper inside the container is installed on the lid of the container, while at the bottom of the container with a gap with respect to a desiccant is placed on the rod, and a fixture for fixing it is installed (SU 1017343, 05.15.1983). However, the design of this container is complex and the absorbent paper used in the design does not allow taking into account the volume of the blood sample, which is important in laboratory analysis.

A device for receiving, storing and transporting dried samples of liquids is known, containing a protective container in which an adsorption element is arranged, which is capable of applying aliquots of liquid to it, analyzed components. The adsorption element is made of a moisture-absorbing porous material with an adsorption capacity of at least 40 mg of water per cm ² capable of reversible desorption of dry components in a sample solvent by immersing at least the working area of the adsorption element with a dried sample in a sample solvent with efficiency not less than 30% after not more than 300 s after the contact of the dried sample with the sample solvent. The device allows to obtain reproducible results in the analysis of biological fluids, and also provides a simplification of the design of the sorbent element (RU 2519030, 10.06.2014). The disadvantages of this device are the complexity in its design and narrow scope, as well as the limited rate of absorption of biofluids.

Also known is the rather simple collection and transportation of biological materials on paper, which has been used for several decades to transport, for example, blood for screening newborns. There is a standard protocol used to accomplish this (Therrel, B. L., ed., Laboratory Methods for Neonatal Screening, American Public Health Association, Washington D.C., 1993). There is also standard paper used for collecting blood samples, which is commonly used.

In particular, an absorbent paper sheet (filter paper) is known having a plurality of area samples, each circle-shaped area sample having a perforation extending substantially along its entire perimeter so as to prevent the capillary liquid sample from flowing from one zone to another. In addition, the absorbent paper is divided into pieces by barrier perforations located between adjacent areas of the samples, in order to provide a gap between the zones of the samples and create an additional capillary flow barrier. The absorbent paper element can be adapted to store any desired sample in liquid form, for example, blood samples or its products, samples can also be arranged in a row for monitoring and calibration purposes, in accordance with known practice (US 5516487, May 14, 1996). The disadvantages of the absorbent paper sheet are the impossibility of accurately dosing the amount of applied sample, the difficulty in removing the circles of the membrane with the sample for their subsequent desorption and its fragility.

A device for collecting and storing biological materials is also known, including a card holder having a window, as well as a fiberglass membrane placed in the window for uniformly absorbing a sample of a drop of blood into a uniform round spot, followed by drying. A fiberglass membrane is used to provide better analysis for small molecules using liquid chromatography / mass spectrometry. The card carrier has a nominal size of approximately 2 inches (5.08 cm) by 3 inches (7.62 cm), and can be made of chipboard. A window made in the card allows the user to apply blood stains directly to the fiberglass membrane without removing it from the carrier. The blood stain is then dried, then knocked out of the fiber membrane and extracted in the laboratory for analysis of small molecules such as pharmaceutical compounds and narcotic substances, etc. (US 2012045792, 02.23.2012). However, this device has a narrow scope, complex design and has no data on the possibility of long-term storage and transportation of the bio-sample. However, it does not have sufficient sterility and integrity properties, since the membrane with the biomaterial applied to it is open to the environment. This source of information is accepted by us as the closest analogue.

The technical result of the claimed invention is to simplify the design of the device while expanding the scope of its application due to the use in the design of the sorption membrane not only for liquid biological materials of living organisms, but also plants and to increase the sensitivity of laboratory research methods, by increasing the amount of absorbed liquid and the possibility to preserve its integrity and isolation from contact with anything, if the transport card is available, as well as to expand the arsenal means for the collection, storage and transportation of biological materials of living organisms and plants.

The technical result is achieved by the fact that a device was created for collecting, storing and transporting biological materials of living organisms and plants, containing a carrier card with sorption material placed therein, configured to deposit biological material containing analyzed components on it, and then drying it, while the device contains a transport card made of coated paper with a density of 300 g / m ² and a stiffness (MD) of 15 mNm in the form of a rectangle with two vertical folds and with the possibility of folding it into three layers, while in the lower layer formed after folding the rectangle along two vertical folds, a sorption membrane made of a moisture-absorbing porous material with a liquid absorption rate of at least 1 cm / s and capillary is rigidly fixed lifting not less than 20 mm / min.

In a preferred embodiment, the drying of liquid biological samples deposited on the sorption membrane occurs within 1.5 hours.

In another preferred embodiment, the transport card, with a sorption membrane located therein, is treated with ultraviolet light at a wavelength of 254 nm for at least 30 minutes to maintain the sterility of the biomaterial.

In a preferred embodiment, the sorption membrane is also capable of reversibly desorbing the dry components in water or another solvent for the sample with an efficiency of at least 80% for 10 minutes after the dried sample contacts the sample solvent.

In a preferred embodiment, the sorption membrane absorbs at least 150 μl of liquid, which allows to increase the sensitivity of laboratory tests.

In another preferred embodiment, the device is configured to analyze not only the liquid biological materials of living organisms, but also plants.

In a preferred embodiment, the sorption membrane also has high tensile strength, which guarantees the integrity of the membrane during transportation and storage to the purpose of its use in the laboratory.

In a preferred embodiment, the sorption membrane is configured to separate pieces with a predetermined area.

Also in a preferred embodiment, the transport card provides isolation of the sorption membrane from the environment and maintains its integrity and sterility.

In addition, in a preferred embodiment, the device is the object of storage and transportation of biosamples to laboratories by any available means of transport (mail, courier, car or air, etc.).

A brief description of the drawings.

In FIG. 1 shows a general view of a transport card containing a sorption membrane integrated therein.

In FIG. 2 shows the outer part of the back layer of the transport card, on which information for the user is printed.

The claimed device for collecting, storing and transporting biological materials of a living organism and plants contains (see Fig. 1 and 2) a transport card (1) made of coated paper with a density of 300 g / m ² and a stiffness (MD) of 15 mNm, which provide paper ability to withstand bending and tearing. The transport card (1) has the shape of a rectangle, the vertical line of which has two folds (2), which make it possible to add it into three layers (3, 4, 5). In the lower inner layer (4), formed after the rectangular transport card (1) is folded along two vertical folds (2), a sorption membrane (6) is made rigidly made of a moisture-absorbing porous material with a liquid absorption rate of at least 1 cm / s and capillary lifting not less than 20 mm / min. The sorption membrane (6) is made with the possibility of applying biological material on it containing the analyzed components, while it preferably absorbs at least 150 μl of liquid, which makes it possible to increase the sensitivity of laboratory tests. In addition, it has a high tensile strength of at least N / 17 mm, which guarantees the integrity of the membrane during transportation and storage to the purpose of its use in the laboratory and the possibility of separating pieces with a predetermined area, which increases the accuracy of laboratory analysis. Prior to use, to preserve the sterility of the biomaterial, a transport card with a sorption membrane placed in it is treated with ultraviolet light at a wavelength of 254 nm for at least 30 minutes. This allows the consumer to supply a quality sterile product. After applying biological material to the sorption membrane (6), it is dried for at least 1.5 hours at room temperature. The sorption membrane (6) is also capable of reversibly desorbing the dry components in water or another solvent for the sample with an efficiency of at least 80% for 10 minutes after the contact of the dried sample with the sample solvent. The transport card (1) provides isolation of the sorption membrane (6) from the environment and preserves its integrity and sterility. In addition, the device is capable of analyzing not only liquid biological materials of living organisms, but also plants, and it is an object of storage and transportation of biosamples to laboratories by any available means of transport (mail, courier, car or air, and other carriers).

Examples of the device.

Example 1

To protect the sorption membrane, which will contain biological material for research, a transport card was made from coated paper with a density of 300 g / m ² and a stiffness (MD) of 15 mNm, which give the paper the ability to withstand bending stress and tearing. The transport map is made in the form of a rectangle folded in three layers due to the presence of two folds on the rectangle. A sorption membrane is glued into the lower inner layer. Protective element - the transport card is arranged so that the sorption membrane is closed from 4 sides, which allows it to maintain its integrity and isolates it from contact with anything. After manufacturing, a transport card with a sorption membrane located in it is treated with UV at a wavelength of 254 nm for 30 minutes. The UV-treated transport card is placed in a plastic bag with a Zip Lock, which previously passed the dry sterilization stage, which allows the consumer to deliver a high-quality sterile product and at the same time close the transport card from external damage. The transport card in the package can be sent by courier, sent by mail and other carriers to the laboratory for its direct use - applying biomaterial to it, drying it, and then, if necessary, storing and transporting it to another laboratory for analysis of the dried biomaterial.

Example 2

To determine the capillary rise, the tip of the sorption membrane was carefully immersed vertically 1-2 mm in water and the distance was determined at which the water front extended for 1 minute, the value of which was at least 2 cm. The specific gravity of the sorption membrane was 152 g / m ² , which was determined by weighing the test sample.

Example 3

A biomaterial mixed with pre-calculated cloned plasmid structures until completely absorbed was applied to a sorption membrane placed in a transport card and pretreated with UV at a wavelength of 254 nm for 30 min. The membrane was dried at room temperature from 20 ° C to 25 ° C for 1.5 hours. The dried sorption membranes were cut with scissors along the lines drawn on them with a known area. The lines help to accurately cut the sorption membrane into pieces of the right size so that the pieces fit into a 1.5 ml tube and are completely immersed in 300 μl of a solution or water that are involved in the elution process. Elution occurred for 10 min with periodic vortexing at room temperature. Then it was centrifuged for 20 s at 10 thousand rpm. The resulting liquid was transferred to a clean tube and used in quantitative PCR. Quantitative analysis of PCR allowed us to calculate the cloned plasmid structures, comparing the initial applied amount on the membrane with eluted from it. The results shown in table 1 showed that the initial number of plasmid structures from the final one differed by 15-18%. This allows us to conclude that the transfer of dry material into the liquid phase occurs with an efficiency of at least 80% within 10 min after the contact of the dried sample with the solvent.

Example 4

To check the degree of liquid absorption of the described sorption membrane, a larger membrane was prepared than the sorption membrane from the transport card. Inside the common sorption membrane, the area corresponding to the sorption membrane from the transport card was drawn with a pencil. In the center, 150 μl of tinted water was added to it. Visually observed the distribution of fluid across the membrane. No transfusion of fluid over the drawn boundaries was observed. In the process of visual observation, it was found that places not filled with water remained on the sorption membrane. This suggests that the sorption membrane, packed in a transport card, allows you to absorb at least 150 μl of liquid. Standard methods for the isolation of DNA and RNA use 100 μl. Therefore, the amount of liquid deposited on the sorption membrane is sufficient to conduct a study with high sensitivity.

Example 5

Transport cards were placed with the lower outer layer on a solid surface (table), the third and fifth layers were carefully opened so that the sorption membranes were freely accessible and 1 cm from the inner lower layer of the transport card. Then they applied biological samples of humans and animals (blood, urogenital smears), as well as freshly squeezed plant juice, selected mechanically (squeezing). Samples on the sorption membrane were dried at room temperature for 1.5 hours. Then the sorption membrane was covered with layers, forming a transport card, which in turn was placed in a plastic bag with a lock and a desiccant. Some of the transport cards were sent for research to a third-party organization, through a transport company. The transport card was tested for tearing using ASTM D 828-87, “Standard Method for Determining the Tensile Strength of Paper and Cardboard under Tensile Strength.” After checking, the paper has excellent mechanical properties on the surface of the paper. The transport card and sorption membrane, when transported to the city located 800 km from the place of packaging and application of the images, they retained their integrity, and the other part of the transport cards with biosamples were stored in a household refrigerator for a month. and the sorption membranes were aliquoted. A liquid aliquot was examined on the same day.

After a month, elution of the dried material from the sorption membrane was carried out. To do this, using scissors, the carrier was carefully cut along the lines dividing it into previously known areas, allowing the sorption membrane to fit into a 1.5 ml tube containing 300 μl of solution or water. The tube was left at room temperature for 10 minutes, periodically shaking on a vortex. Then the tube was centrifuged, the supernatant was taken and transferred to a clean tube. The obtained liquid sample was used in further work for DNA isolation and subsequent PCR. The results of studies of liquid and dry samples are presented in table 2.

Example 6

The sterility of the transport card and sorption membrane was checked by bacteriological seeding. For this, the sorption membrane was immersed in bacteriological meat broth and agar. Then they were placed at 37 ° C for three days. After three days, the broth and agar were checked for bacterial outgrowths. No seedlings were found. Sterilization of the transport card and the UV membrane at a length of 254 nm for 30 minutes and a plastic bag by dry sterilization allows you to have a sterile product for its intended use.

Thus, the claimed invention is compact, convenient to use, provides both high sensitivity in conducting laboratory analysis, and good preservation and transportation of the bio-sample over large and short distances for a long time.

Claims (5)

1. A device for collecting, storing and transporting biological materials of living organisms and plants, containing a carrier card with sorption material placed therein, made with the possibility of applying biological material containing analyzed components on it and then drying it, characterized in that as a carrier card, the device contains a card made of coated paper with a density of 300 g / m ² and a stiffness (MD) of 15 mNm in the form of a rectangle with two vertical folds and with the possibility of folding it in three layers, while in the lower inner layer formed after the rectangle is folded over two vertical folds, a sorption membrane is made rigidly made of a moisture-absorbing porous material that absorbs at least 150 μl of liquid with a liquid absorption rate of at least 1 cm / s and capillary lift of at least 20 mm / min. 2. The device according to claim 1, characterized in that the carrier card, with the sorption membrane located in it, is treated with ultraviolet light at a wavelength of 254 nm for at least 30 minutes. 3. The device according to p. 1, characterized in that the sorption membrane is capable of reversible desorption of dry components in water or another solvent for the sample with an efficiency of at least 80% for 10 minutes after contact of the dried sample with the solvent for the sample. 4. The device according to p. 1, characterized in that it is configured to analyze not only liquid biological materials of living organisms, but also plants. 5. The device according to p. 1, characterized in that the sorption membrane is configured to separate pieces with a predetermined area.

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