CN102608312A

CN102608312A - Blood serum or blood plasma diluting solution applied to detection method of trace gelatin particle agglutinated HIV-1 (Human Immunodeficiency Virus-1) recent infection

Info

Publication number: CN102608312A
Application number: CN201210046148XA
Authority: CN
Inventors: 李洪
Original assignee: Individual
Current assignee: Individual
Priority date: 2012-02-28
Filing date: 2012-02-28
Publication date: 2012-07-25

Abstract

The invention provides a blood serum or blood plasma sample diluting solution applied to a method for detecting trace gelatin particle agglutinated HIV-1 (Human Immunodeficiency Virus-1) recent infection, belonging to the field of a biological technology. The invention particularly provides a reagent applied to detecting the HIV-1 recent infection. The diluting solution is prepared from a buffering solution, TritonX100 with the concentration of 30%, NaN3 and bovine serum albumin by mixing according to a ratio. The diluting solution disclosed by the invention can completely replace a DilsimTM11 diluting solution and can be used for a detection method of the trace gelatin particle agglutinated HIV-1 recent infection.

Description

A kind of serum or plasma sample diluting liquid that is applied to micro-gelatin particle aggegation HIV-1 recent infection detection method

Technical field

The invention belongs to biological technical field, the reagent of using in the especially a kind of HIV-1 of detection recent infection.

Background technology

AIDS is a kind of infectious disease, and detecting AIDS virus (HIV) is the important method of finding the infected.In order to grasp its fashion trend, estimate the effect of prevention and control, countries in the world are all wanted to use HIV New Development infection rate and are estimated, and the calculating of New Development infection rate then needs HIV recent infection data and carries out.

HIV recent infection data can detect HIV recent infection method through the kinds of experiments chamber and obtain.Whether HIV recent infection method is detected in the laboratory can be that serum changes before the sun or serum changes sun and classified afterwards according to the sample that uses.The method that serum changes the sample employing before the sun is to detect P24 antigen or the HIV-1 RNA of HIV-1 in the sample; The method that serum changes the sample employing behind the sun has methods such as 3A11-LS, BED-CEIA, and they belong to serology and detect a type (STARHS) that recent HIV-1 serum changes positive method.

Except that above-mentioned method, nearly 20 kinds of the laboratory detection method of being set up till now.Trace gelatin particle aggegation HIV-1 recent infection detection method, promptly ML-PA method (name of using in the past is called PA-LS) is to be formed by a kind of HIV of primary dcreening operation simply and easily gelatin particle aggegation method (PA) improvement of generally acknowledging in the world at present.The PA method is to carry out with the HIV AIDS diagnosis reagent that Japanese Fujirebio company produces; It is through being coated on HIV-1 antigen (HIV-1/gp41 and HIV-1/P24) on artificial carrier's gelatine particle; HIV-1 antibody in sensitization ion and the blood reacts the generation aggegation; The reaction of generation particles aggregate can detect the HIV-1 antibody in serum and the blood plasma thus.

BED HIV-1 recent infection detection method compares the characteristics that have more simply, cheaply, do not need instrument and equipment and be highly susceptible to promoting to trace gelatin particle aggegation HIV-1 recent infection detection method with using comparatively widely at present in the world.Its price is merely 1/20th to a thirtieth of BED method.But because the dilution of serum that uses in this method or plasma sample is BioMerieux (Raleigh, NC) Dilsim of the Vironostika HIV antibody dedicated test reagent of company's production ^TMTherefore the ll dilution costs an arm and a leg, and effectively storage life is very short, can not provide separately generally speaking, can't buy in China to obtain.Thereby, limited of the widespread use of this method greatly in China.

Summary of the invention

The dilution that just provides a kind of serum or plasma sample to be solved by this invention is for micro-gelatin particle aggegation HIV-1 recent infection detection method (ML-PA) can be in the problem of China's widespread use.This dilution can substitute the Dilsim that uses in the ML-PA detection method ^TMThe ll dilution, and do not influence the experiment and the testing result of this method.

Serum or the plasma sample diluting liquid that is applied to micro-gelatin particle aggegation HIV-1 recent infection detection method of the present invention; It is characterized in that this dilution is mixed with by damping fluid, 30%Triton X 100, NaN3 and bovine serum albumin(BSA) in proportion; Wherein, damping fluid is by Na ₂HPO ₄, Na ₂HPO ₄Be dissolved in the deionized water with NaCl and be prepared into, add 1.27 gram Na in per 1000 ml deionized water respectively ₂HPO ₄, 0.3 gram Na ₂HPO ₄With 0.85 gram NaCl; This dilution is damping fluid by weight: NaN3: bovine serum albumin(BSA)=1002.42:0.2:10, by volume 30%Triton X 10: deionized water=1:100.

Wherein, damping fluid helps the pH value that keeps constant, is beneficial to the abundant reaction of antigen-antibody.30%Triton X 100 is a kind of comparatively gentle scaling agent, and surfactant or title interfacial agent, Chang Zuowei adjuvant make albumen keep stable, makes the albumen in serum or the plasma sample keep stable.NaN3 is as serum plasma sample antiseptic.Bovine serum albumin(BSA) is as the protective agent of serum plasma sample antibody.

Serum of the present invention or plasma sample diluting liquid; Be used for micro-gelatin particle aggegation HIV-1 recent infection detection method; Dilution can be realized autonomous production; Can be at home generally and widely use, can become China and formulate effective policy, carry out the action of effective reply AIDS, even formulate relevant policies and planning provides effective technical support for the various countries decision maker.

Dilution of the present invention is applied to micro-gelatin particle aggegation HIV-1 recent infection detection method (ML-PA) and adopts BioMerieux (Raleigh, NC) special-purpose serum of company or plasma sample Dilsim ^TMThe PA-LS method of ll dilution is compared and is had following difference and superiority:

Have following characteristics and superiority and compare with Vironostika-LS that generally adopts and BED-CEIA method with the ML-PA method of dilution of the present invention:

Embodiment

Embodiment 1: a kind of serum or plasma sample diluting liquid that is applied to the detection method of micro-gelatin particle aggegation HIV-1 recent infection, this dilution is by 1.27 gram Na ₂HPO ₄, 0.3 gram NaH ₂PO ₄0.85 being dissolved in, gram NaCl adds 10 milliliters of 100,200 milligrams of antiseptic NaN3 of 30%Triton X that increase permeability of cell membrane in the damping fluid that 1000 ml deionized water are prepared into; And the protective agent bovine serum albumin(BSA) of 10 gram antibody, fully mix acquisition.

In order to confirm dilution of the present invention and Dilsim ^TMThe consistance of ll dilution with 450 H IV positive serum samples through the unknown infection time of affirmation, is carried out the test that recent infection detects.Each sample comes with dilution of the present invention and Dilsim by 6 gradients of 1: 10,1: 100,1: 1 000,1: 10 000,1: 20 000,1: 40 000 ^TMThe ll dilution dilutes, and for three days on end, carries out 3 tests in same laboratory by two different people once a day.Detected recent infection sample is 27 parts, and two kinds of detected recent infection samples of dilution are in full accord, and the dilution gradient result of each sample is duplicate, explains that dilution of the present invention can substitute Dilsim fully ^TMThe ll dilution is used for the detection method of micro-gelatin particle aggegation HIV-1 recent infection.

Claims

1. a serum or plasma sample diluting liquid that is applied to micro-gelatin particle aggegation HIV-1 recent infection detection method; It is characterized in that this dilution is mixed with by damping fluid, 30%Triton X 100, NaN3 and bovine serum albumin(BSA) in proportion; Wherein, damping fluid is by Na ₂HPO ₄, Na ₂HPO ₄Be dissolved in the deionized water with NaCl and be prepared into, add 1.27 gram Na in per 1000 ml deionized water respectively ₂HPO ₄, 0.3 gram Na ₂HPO ₄With 0.85 gram NaCl; This dilution is damping fluid by weight: NaN3: bovine serum albumin(BSA)=1002.42:0.2:10, by volume 30%Triton X 10: deionized water=1:100.