CN103571914A - Acquisition method and device for obtaining nucleic acid clinical specimen - Google Patents

Abstract

The invention relates to an acquisition method and an acquisition device for obtaining a nucleic acid clinical specimen. Air exhaled by a patient is collected through the collection device and is filtered through a filtration membrane of the collection device; the filtration membrane is immersed in TE (Tris/Ethylenediamine Tetraacetate) or de-ionized water; the collection on the filtration membrane is dissolved by using ultrasound; dissolved liquid is collected, wherein the dissolved liquid is the nucleic acid clinical specimen.

Description

Collection method and device for obtaining nucleic acid clinical specimens

technical field

The invention relates to a collection method and device for obtaining nucleic acid clinical specimens.

Background technique

During medical testing, it is often necessary to collect nucleic acid clinical specimens from patients. Most of the existing collection methods are to collect 3-5 ml of peripheral blood or 1-2 mg of tissue samples from patients, and extract nucleic acids through kits to obtain nucleic acid clinical samples. However, there is no good method to collect and extract free nucleic acid samples from patients' exhaled breath.

Contents of the invention

The object of the present invention is to provide a collection method for obtaining nucleic acid clinical specimens, which does not require blood collection, is easy to operate, and has good collection effects.

Based on the same inventive concept, the present invention has two independent technical solutions:

1. A collection method for obtaining nucleic acid clinical specimens, characterized in that: the air exhaled by the patient is collected by the collection device, the air exhaled by the patient is filtered by the filter membrane of the collection device, and the filter membrane is soaked in TE buffer solution or deionized water , using ultrasound to dissolve the collected matter on the filter membrane, and collect the eluate, which is a clinical specimen of nucleic acid.

When soaking the filter membrane in TE buffer or deionized water, centrifuge at 1000g for 10 minutes.

When collecting the eluate, it needs to be centrifuged at 13000g for 10 minutes.

Collect 5-10L of air exhaled by the patient through the collection device.

2. The acquisition device for realizing the above method is characterized in that: the air collection tube, the filter tube and the eluate collection tube are plugged sequentially from top to bottom, the filter tube is provided with a filter membrane, and the bottom end of the filter tube is provided with an opening, A closing cover is provided at the opening, and a tube cover is provided at the top of the filter tube.

Both the upper part of the air collecting pipe and the lower part of the filter pipe are tapered.

The opening at the bottom end of the filter tube is threadedly connected with the closure cap.

The beneficial effect that the present invention has:

Microorganisms or the nucleic acid substances they release form aerosols in the air, and when the aerosol passes through the filter membrane, the macromolecular nucleic acids in it are trapped on the filter membrane. Soak the filter membrane with deionized water or TE buffer solution, dissolve the collection on the filter membrane by ultrasound, and collect the eluate to obtain the nucleic acid clinical specimen. The present invention does not require blood collection, and only needs to be exhaled by the patient to realize the collection of nucleic acid clinical specimens. The operation is simple, fast, efficient, and non-invasive. The collection device of the present invention can not only be applied to medical detection such as tuberculosis detection, but also can be widely used in environmental monitoring.

Description of drawings

Fig. 1 is the structural representation of collecting device of the present invention;

Fig. 2 is a schematic structural view of the filter tube of the collection device of the present invention.

Detailed ways

As shown in Figure 1, the collection device includes an air collection tube 1, a filter tube 2, and an eluate collection tube 3, and the air collection tube 1, filter tube 2, and eluate collection tube 3 are inserted in sequence from top to bottom, and the bottom of the filter tube is Leave a space between the end and the bottom of the eluate collection tube. The air collection tube 1 is used to receive the gas exhaled by the patient, and can also be connected with other gas supply equipment. The upper part of the air collection pipe 1 and the lower part of the filter pipe 2 are both tapered. As shown in Figure 2, a filter membrane 5 is arranged inside the filter tube, and the filter membrane is made of glass fiber, silicon crystal fiber, TCLP or silica gel. The filter membrane 5 is placed on the bracket 6, and the bracket 6 has a mesh structure, which supports the filter membrane 5 and prevents the filter membrane 5 from falling off. The top of filter tube is provided with cap 4, and the bottom of filter tube is provided with opening, and opening is provided with closure cap 7, and the outside of the opening of filter tube bottom is provided with external thread, and the inside of closure cap is provided with internal thread, and the bottom of filter tube The opening is threadedly connected with the closing cap, and the opening at the bottom end of the filter tube can also be connected with equipment such as a negative pressure guide.

When collecting nucleic acid specimens, connect the air collection tube 1 and filter tube 2, and do not use the closing cap 7 at this time; after the patient inhales deeply, close the mouth of the patient with the air collection tube, and the patient exhales forcefully, repeat 3 times, and collect 5-10L patient exhales; then tighten the closure cap 7, remove the air collection tube 1, insert the filter tube 2 into the eluate collection tube; add 800 microliters of TE buffer or deionized water to the filter tube, cover the tube Cover, and centrifuge at 1000g for 10 minutes to make the liquid completely infiltrate the filter membrane 5; open the tube cover 4, insert an ultrasonic probe, and dissolve the collection on the filter membrane 5 by ultrasonication for a short time; unscrew the sealing cover 7, cover the tube cover 4, Put the filter tube into a new eluate collection tube; centrifuge at a high speed of 13000g for 10 minutes, the eluate collected in the eluate collection tube is the nucleic acid sample to be tested, and store it at low temperature.

Claims (8)

1. an acquisition method that obtains nucleic acid clinical samples, it is characterized in that: by gathering device, gather the air that patient breathes out, the air filtration of patient being breathed out by the filter membrane of gathering device, filter membrane is soaked in TE damping fluid or deionized water, utilize ultrasonic the gleanings stripping on filter membrane, collect dissolution fluid, dissolution fluid is nucleic acid clinical samples.

2. the acquisition method that obtains nucleic acid clinical samples according to claim 1, is characterized in that: when filter membrane is soaked in TE damping fluid or deionized water, need 1000g centrifugal 10 minutes.

3. the acquisition method that obtains nucleic acid clinical samples according to claim 2, is characterized in that: while collecting dissolution fluid, need 13000g centrifugal 10 minutes.

4. the acquisition method that obtains nucleic acid clinical samples according to claim 3, is characterized in that: by gathering device, gather the air 5-10L that patient breathes out.

5. the acquisition method that obtains nucleic acid clinical samples according to claim 4, is characterized in that: filter membrane is made by glass fibre, silicon fibre, TCLP or silica gel.

6. one kind for realizing the gathering device of method described in claim 1, it is characterized in that: air trapping pipe, strainer tube and dissolution fluid collection tube are from top to bottom pegged graft successively, in strainer tube, be provided with filter membrane, the bottom of strainer tube is provided with opening, opening part is provided with closing cap, and the top of strainer tube is provided with pipe lid.

7. gathering device according to claim 6, is characterized in that: the top of air trapping pipe, the bottom of strainer tube are all tapered.

8. gathering device according to claim 7, is characterized in that: the opening of strainer tube bottom is threaded with closing cap.

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