wqrj posted an update //www.wtai.cn/ ( a ) antibody detection are mainly enzyme-linked immunosorbent assay ( ELISA ) and immunofluorescence test ( IFA ) .ELISA detergent cracking HIV or infected cells extract antigen ,IFA infected cell smears as antigens for antibody testing ,if found positive specimens should be repeated once .
In order to prevent the false positive ,can be Westernblot ( WB ,Western blotting ) further confirmation .The WB method is using polyacrylamide gel electrophoresis of HIV proteins were separated by electrophoresis ,and then transferred to different protein band transfer to nitrocellulose membrane ,joining the patient serum after incubation, using antihuman globulin protease labelled antibody staining ,can measure according to the different structure of protein ,such as resistance to GP12 ,gp41 ,P24 antibodies ,specific high .
Rapid detection method, also known as the gold standard ,and antibody detection method ,based on the principle of immunity chromatography assay for detecting HIV antibody ,qualitative detection .
( two) antigen detection using ELISA P24 antigen detection ,early in HIV infection has not yet appeared antibodies ,blood is the presence of the antigen .Because P24 amount is too small,the positive rates are usually low .
Existing dissociation of immune complexes or condensed P24 antigen,to increase sensitivity .( three) nucleic acid detection using PCR assay for detection of HIV gene ,is fast ,efficient ,sensitive and specific method, the method has been applied to HIV for early diagnosis of HIV infection and AIDS research in .
( four) virus isolation methods for co-culture method ,namely the use of normal human peripheral blood mononuclear cells isolated and cultured with PHA stimulation ,after joining the patient ,mononuclear cell diagnosis and AIDS research in .
HIV laboratory techniques and methods Key words acquired immunodeficiency syndrome HIV laboratory techniques and methods of AIDS ( AIDS ) also known as acquired immunodeficiency syndrome ,is by the human immunodeficiency virus ( HIV ) infection caused by an immune deficiency disease .
HIV diagnosis of infection is the AIDS prevention and control work important component ,the establishment of sensitive and practical detection method for monitoring ,diagnostic or screening blood ,control the epidemic of AIDS is very important .
Following the HIV detection technology and progress of .HIV infection mechanism of HIV virus invades the human body, its surface glycoprotein GP12 andcell surface receptor protein CD4 with high affinity binding ,adsorption to the host cells ;GP12 and cell surfaceauxiliary receptor interactions ,allowing the virus and host cell membranes closer ;gp41 produced a series of conformational changes ,the N end of the fusion peptide fragment is inserted into the host cell membrane, leading to viral envelope with the cell membrane of the final fusion ,RNA virus into the cell .
In the HIV after infection, the first to detect the virus RNA ,then p24 antigen,finally the .HIV infection markers in infection after 1~ 14 d,RNA virus level is rising exponentially ,followed by a decline and maintained in a stable level, enter HIV period of asymptomatic .
P24 antigen levels with the viral RNA levels of development, during the acute phase of infection can occur ,is considered to be the indirect markers of viral replication ,but because the detection sensitivity is not enough and the detection time than the RNA late .
From HIV infection to detect antibodies to HIV during this period, known as the window period .In the window period, through the RNA virus ,p24 antigen and CD4lymphocyte levelsto determine the HIV infection .
CD4 lymphocyte levels with infection develops gradually decreased, when the blood cells decreased to 2/mL,can occur in patients with severe immunodeficiency ,was diagnosed with aids .Virus RNA ,p24 ,HIV and CD4 lymphocyte antigen antibody levels can be used to determine HIV infection ,detect the condition development .
HIV in viral classification HIV in viral classification in the genus Retroviridae lentivirus in human immunodeficiency virus group .To date ,based on serologic responses and viral nucleic acid sequence determination ,the global epidemic of HIV can be divided into 2 types:HIV ?1and HIV?2.
In the HIV ?1 type,according to the coding of envelope protein env gene and coding protein gag gene sequence homology is further divided into 3 groups: group M( primary group) ,group O ( peripheral group) and group N ( new group or non M non O group ) ,M group and can be divided into A ?J1 subtypes.
In the HIV ?1and HIV?2,its nucleotide sequence homology was 45% ,and in the presence of immune cross reaction .Immunoblot assay ( West blot ) two can be clearly separated .The primary HIV infection detection method is a method for detecting HIV 1,generally can be divided into antibody detection and virus detection in two categories .
Virus detection including cell culture ( virus ) ,p24 antigen detection and virus nucleic acid testing .Early diagnosis of HIV is mainly by serological tests for the detection of anti HIV antibody ,indirect diagnosis of HIV infection .
In recent years ,the molecular biology methods have been applied to the detection of HIV ,HIV laboratory diagnostic methods have made great progress ,nucleic acid detection has become the development direction of HIV laboratory diagnosis .
Antibodies are usually 3to 8 weeksafter infection can be detected .At present, most of the double antigen sandwich method ,this method has a good sensitivity and specificity .Antibody detection by screening and confirmation tests ,screening tests must be performed to confirm the test positive .
Enzyme linked immunosorbent assay ( ELISA ) method has certain sensitivity ,and has the advantages of simple operation ,rapid ,suitable for a large number of sample testing .Therefore ,universal screening is currently in clinical detection method .
International has 3 confirmed test ,including Western blot test ,strip immunoassay and immunofluorescence test ,the most commonly used in Western blot test .In the window period, HIV antibody can be detected ,but can detect the virus antigen or isolation of virus .
Antigen in individual infection before seroconversion after 2～ 18 Dwas detected .Therefore ,in serum transforming period through detection of p24 antigen has great advantages ,can be used as the early diagnosis of HIV infection as a method of .
Viral culture of virus culture is the most accurate method for detection of HIV infection .General to take the culture of peripheral blood mononuclear cells ( PBMC ) method for the diagnosis of HIV .
Detection of virus nucleic acid is usually through detection of HIV RNA level to reflect the viral load ,with very high sensitivity, using a real-time polymerase chain reaction ( PCR ) technology ,can be in HIV infection 2 weeks prior todetect viral nucleic acid .
Virus nucleic acid testing method can be used for early diagnosis of HIV ,such as window period, diagnosis, course ,guiding treatment and monitoring for efficacy determination ,prediction of disease course .
The common determination method with reverse PCR experiment ( RT ?PCR ) ,nucleic acid sequence-based amplification experiment (NASBA ) ,branched DNA hybridization experiments (bDNA) etc.
.Using the high sensitivity of the real-time PCR technology ,can be in HIV infection 2 weeks prior todetect viral nucleic acid ,.HIV antibody ,antigen for detection of HIV antibody in human infection after several weeks to appear gradually, can be extended to ,serological tests for screening and confirmation tests .
The most commonly used screening test and validation test are ELISA and Western blot test ( WB ) .Conventional experimental methods: ELISA ,immunoblotting test ( WestBlot ) ,indirect immunofluorescence ( IFA ) .
Rapid detection method :gelatin particle agglutination test ,dot immunobinding assay ,P24 antigen detection,molecular biology methods ,RT ?PCR detection ,real time fluorescence PCR detection technology ,branched DNA ( bDNA ) ,ligase chain reaction enzyme ( LCR ) ,nucleic acid sequence dependent amplification ( NASBA ) ,genuine leather briefcases for men wholesale,transcription mediated by amplification ( TMA ) enzyme-linked immunosorbent assay ELISA law is the basic principle of immune reactants through chemical or immunological method for the formation of enzyme conjugates ,enzyme conjugates with the sample to be detected the corresponding antigen or antibody binding to become immune complexes ,then adding enzyme substrate ,the enzyme that catalyzes the hydrolysis of the substrate or ,colorless to produce color ,with the naked eye ,spectrophotometer observation results .
Screening with HIV ELISA reagents have been developed to the fourth generation detection reagent .The first generation of reagents to viral lysates or partially purified virus antigen coating reaction plate ,to the detection of antibody in serum .
As the bag is the antigen is not very pure ,the false positive rate was high .The second generation of reagents using gene engineering method to obtain recombinant antigens and synthetic peptide coated reaction plate ,due to the use of purified antigen ,specificity has been greatly improved .
The third generation of reagents using double antigen sandwich assay for the detection of antibodies ,to further improve the sensitivity of .The fourth generation of reagents in the third generation based on the further increase of the P24 antigen detection,the HIV antigen and antibodies to the P24 packis a reaction plate ,which can simultaneously detect the serum HIV antibody and P24 antigen .
Western blot test immunoblot test is used for validation testing ,is the basic principle of HIV virus antigen by SDS ?PAGE electrophoresis ,wholesale laptop bags,the molecular weight proteins ranging in size with separated, and then put these have different separating proteins transferred to nitrocellulose membrane charged .
The film cut into strips ,each strip on the cellulose nitrate film containing an electrophoretic separation of the HIV virus antigen .The tested serum samples using a dilute 1/1,then it is added directly to the cellulose nitrate film, constant temperature concussion ,make its full contact reaction ,serum containing antibodies against HIV ,will and membrane on the antigen combining .
Anti human IgG enzyme conjugates and the substrate, to make a response to the antigen antibody binding strip presents ?Purple brown ,judge according to the appearance of strip .The report says ,immunoblot assay specificity is not very good ,there are approximately 2% and the false positive rate ,but the Western blot test is still the most commonly used HIV confirmation test .
Immunofluorescence test ( IFA ) basic principles for application of H ?9 or HUT?78cell culture as acarrier ,with HIV infected cells ,the cells will contain HIV antigen ,HIV infection of lymphocytes is coated on the glass slide, fixed ,preparation for the antigen ,adding serum to be tested ,the serum .
Anti HIV antibody and antigen binding ,and fluorescein labeled anti human Ig binding ,in fluorescence microscopy can be seen within the cell is yellowish green fluorescence .Gelatin particle agglutination test ( PA ) PA is the basic process of first sample dilution ,and adding the antigen sensitized and non-sensitized gelatin particle ,after mixing thermal insulation (usually at room temperature) .
When the serum HIV antibody ,the antigen sensitized gelatin particles and antibody antigen antibody reaction ,based on gelatin particles in Kong Zhong interpretation of results of agglutination .
PA has the advantages of simple operation ,no need of special equipment ,suitable for small samples .Dot blot test ( or immune chromatography / or percolation ) dot blot test using nitrocellulose membrane as solid carrier ,with HIV antigen coating on the solid phase carrier ,adding tested samples ( serum ,plasma ,for urine and other body fluids) ,the temperature response after eluting uncombined on solid carrier specimens ,package antigen to be detected and the serum antibody binding ,using colloidal gold ( or colloidal selenium ) instead of substrate connection in staphylococcal protein A ,gold labeled protein A with human Ig binding ability ,thus can be captured HIV antibody .
If a sample of HIV antibody ,the film will generate a red-orange spots ( or lines ) ,general 3 ～ 1 minresults ,better specificity ,more suitable for remote areas of clinical blood test ,but not suitable for city regions in the screening of blood donors .
Molecular biology method with the development of biological technology ,nucleic acid detection has been more and more attention ,it can directly check the HIV RNA ,can be found in serological changes prior to the detection of HIV infection ,and the ratio of P24 antigen detection method is more sensitive .
Polymerase chain reaction detection method of basic principle of PCR technology DNA similar to the natural reproduction process ,http://www.epssogroup.com/,its specificity depends on the target sequences are complementary oligonucleotide primer .
PCR from degeneration ,annealing ,extension three elementary reaction steps .① templateDNA denaturation :template DNA upon heating to 93 ℃after a certain period of time,so that the template of DNA double-strand or amplified by PCR formation of double-stranded DNA dissociation ,make become single-stranded ,so that it and the primer binding ,as to the reaction to prepare .
The template DNA and primer annealing ( complex ) :template DNA degeneration into single after heated ,the temperature dropped to 55 ℃,primer and template DNA strand complementary sequence pair combination .
③ primerextension: DNA template and primer binding compounds in TaqDNA polymerase function, taking dNTP as reaction raw materials ,the target sequence as a template ,by base pairing with semiconservative replication principle ,the synthesis of 1 newtemplates and DNA chain complementary semiconservative replication chain ,repeated cycles of denaturation ,annealing ,extension three ,you can get more semiconservative replication chain ,and this kind of new chain can become the next circular template .
Each completed 1 cycleneeds 2 ～ 4 min,2～ 3 hcan be expanded to gene amplification millions of times .Detection of HIV ?RNA ,need to use reverse transcription reaction RNA transcription of cDNA, then cDNA template for PCR amplification can be, this reaction is called RT pcr .
Real time fluorescent quantitative PCR technology of real-time fluorescence quantitative PCR technology ,is defined in the PCR reaction system with fluorescent moieties, using fluorescence signal real-time monitoring of the entire process of PCR ,the standard curve for unknown template quantitative analysis method .
This technique is based on the principle of the use of fluorophore labeled probe ,5end labeled with fluorescent groups R ,3end marker quenching group Q ,in the absence of PCR amplification ,because the fluorescent moiety and a quenching group space was very close, making the fluorophore is quenched ,no fluorescence ;and when the PCR amplification ,primer with specific fluorescent probes were combined in the template ,fluorescently labeled probes and template combined position located on the downstream primers ,using Taq enzyme 5&prime ;3?Exonuclease activity ,the fluorescent probe hydrolysis ,fluorescent moiety is released, due to the space and quenching group separately, emit fluorescence .
The emitted fluorescence can be detected by fluorescent probe ,side side of amplification ,detection ,so as to realize the real-time detection .The technology is not only achieved a PCR from quantitative to quantitative leap ,and comparison with conventional PCR ,it has strong specificity ,high degree of automation ,effectively solves the problem of PCR pollution characteristics .
The branched DNA assay -- bDNA bDNA is quantitatively detected in the plasma of HIV ?1RNAmethod .BDNA is synthesized with side chains of the DNA fragment ,in each of its side chains can be labeled by excited marker .
The HIV RNA by centrifugation from viral particles released, and then use the capture probe 1capturedto pores .The capture probes 2and viralRNA in another part of the specific binding ,and pre amplification probe binding .
And then the latter amplification probe bDNA probe hybridization .Two sets of target probes respectively with RNA virus pol gene in different regions of the specific binding of .In the micropores are formed in a HIVRNA oligonucleotide complexes ?.
Then add 1 chemiluminescencesubstrate after incubation may amplify chemiluminescence signal .Through the luminous intensity to quantitatively ,because the light intensity and the content of HIVRNA in proportional sample .
BDNA does not exist of amplified product contamination ,it is PCR is a big progress .BDNA has ten to cover the entire genome of the probe ,can easily detect HIV some variants ,but the sensitivity to PCR ,enhanced bDNA sensitivity is a big problem .
Nucleic acid sequence dependent amplification ( nucleic acid sequence based amplification ,NASBA NASBA ) is composed of a pair of primers mediated ,continuous and uniform ,in vitro specific nucleotide sequence of isothermal amplification of RNA .
Reaction at 42 ℃ for 2 h,in the RNA template is about 19 times .The principle of NASBA is the extraction of viral RNA ,joining the AMV reverse transcriptase enzyme ,RNA H ,T7RNA polymerase and primers for .
The reaction of non circular phase and cycle phase :in non circular phase ,I primer and template RNA after annealing at AMV reverse under the action of the enzyme synthesis of cDNA ,RNA:DNA heterozygotes ,and RNaseH degradation of RNA Ⅱ and cDNA,primer annealing ,in the reverse transcriptase of synthesis of second DNA complementary chain .
Double chain DNA in T7RNA polymerase action, the promoter sequence starting and transcription of RNA ,RNA can also be used in reverse transcriptase under reverse transcribed into DNA ,enter the circulation ,the template for a large amplification .
Transcription mediated amplification technology ( transcription mediated amplification ,TMA ) TMA technology principle and NASBA are basically the same, slightly difference is that TMA is the use of MMLV reverse transcriptase and T7 RNA polymerasetwo enzymes ,MMLV reverse transcriptase both reverse transcriptase activity with RNA enzyme H activity .
Reaction at 41.5 ℃ for 1 h,in the RNA template is about 19 times .Ligase chain reaction enzyme ( LCR ) LCR is based on a target molecule dependent oligonucleotide probes are connected to each other by a probe amplification technique ,is the new development of PCR after a more promising in vitro amplification technology .
Its principle is composed of 2 sections of oligonucleotide single-stranded DNA probes and target sequences on hybridization ,when the 2 segment DNAprobe and no mutations in the template faded after the fire ,if the 2 probeis immediately ,genuine leather laptop handbags wholesale,without intermediate nucleotide interval ,can be connected in ligase under ,after connecting new chain and can be used as a template ,guide the next cycle is connected to generate new chain .
If the connecting segment occurring nucleotide base mutation ,is connected to the reaction does not occur ,the amplification reaction termination .The current detection technology insufficient cell culture method for the detection of HIV strong specificity ,no false positive ,to confirm those of antigen / antibody detection in uncertain individual and positive mothers neonatal HIV infection has important significance .
But the need to have a certain number of infected cells can train and by isolation of the virus ,which has poor sensitivity ,long operation time ,operation is complex ,must be in the particular P3 laboratorycan be carried out, and the cost is higher ( every culture needs about 2 ～ 5 dollars) ,not suitable for .
Detection of p24 antigen in viral replication can be started after detected in blood of soluble p24 antigen, but prone to false positives .Therefore ,positive results must be confirmed by the neutralization test ,the results can be used as auxiliary diagnosis based on HIV infection .
HIV ?1p24 antigennegative ,only that in this experiment has no reaction ,cannot exclude infection of HIV .Virus nucleic acid detection method has high sensitivity ,for the monitoring of disease progression ,antiviral effect and drug resistance monitoring is very important .
However ,due to the diversity of HIV gene ,not a set of primers can cover all HIV sequence ,so that the detection sensitivity and restricted ;in addition to existing virus nucleic acid detection method and detection apparatus ,detection reagent or expensive ,complicated operation ,high requirements on the operators ,is difficult in general laboratory ,also not suitable for a large number of patients with rapid detection, also not suitable for broad clinical application .
Therefore ,it is necessary to improve the detection sensitivity ,specificity ,shorten the window period ,but also is simple ,fast and reduce costs has become a HIV detection technology development requirements and direction ,a lot of research is dedicated to finding the virus detection of alternative technology .
HIV detection technology progress in recent years, great progress has been made in HIV detection technology .As the development of ICD p24 antigen assay ( immune ?Complex disassociate ,immune complex dissociation ,ICD ) ,the fourth generation of HIV detection reagent ,ultra sensitive EIA ,linear immune enzyme assay ( lineal immunoenzymatic assay ) ,allowing the detection of HIV infection time significantly ahead of ,.
HIV detection reagent development from 1985 first generation HIVantibody detection reagents available HIV serological detection reagent has been developed to the fourth generation .The fourth generation ELISA agent has the advantages of simultaneous detection of antigen and antibody ,reduce the residual risk of blood screening .
And the third generation of HIV ?1/2reagents,detection time of 4 ～ 5 d,the window period shortens more than 1 weeks in early diagnosis of AIDS ,obviously better than that of the third generation .
But the use of the fourth generation of reagents on HIV testing, still have 2～ 3week window .In addition ,the antigen and antibody simultaneously coated on the reaction plate ,there exists mutual interference may ,affecting the specificity of the immune response .
But due to the second window period, also makes the detection sensitivity affected .Generally speaking ,the use of fourth generation of reagents for the detection of p24 antigen ,its sensitivity ( 2~ 1 pg/mL) is much lower than the detection of antigen and antibody assay ( 3.
5~ 1 pg/mL) .From the methodological perspective, based on the ELISA analysis method ,sensitivity is limited .Relative chemiluminescence and time-resolved fluorescence immunoassay and other more advanced analysis method ,its sensitivity is relatively low .
Therefore ,improve the sensitivity ,specificity ,shorten the window period and rapid HIV detection reagent is the future development trend .Viral detection methods for the development of nucleic acid detection method research progress with the development of molecular biology technology ,nucleic acid detection has been paid more and more attention and the reaction of antigen and antibody of high specificity to combine to form the immune PCR technology .
This technique has strong specificity and high sensitivity ,can be used for a single antigen detection .This promotes the application of PCR technology for the rapid development of HIV nucleic acid detection .
Since 21 years ,COBAS Ampliscreen HIV ?1 Testand 4HIVnucleic acid detection technology through the FDA approval, as in the screening analysis technology to detection of HIV ?1.Recently, real-time fluorescent PCR detection technology to make HIV nucleic acid detection technology into a new .
By this technique ,not only can carry out qualitative detection, the more important is that it can detect .Barletta, this technology will be applied to the detection of HIV ,greatly reduce the viral load detection limit ( .
66 copies ofthe equivalent of.33 virus particles) .In April 22, the State Drug Administration ( SDA ) approved the first HIV fluorescence PCR detection reagent kit .P24 antigen detection method research progress along with the detection of continuously improve the sensitivity ,detection of p24 antigen gradually from is mainly used in the window period assist early diagnosis and further shortening of the window period ,development for viral load determination .
At present, the p24 antigen in the following aspects have important applications :HIV ?1antibodies do notdetermine or the window period of diagnosis ;HIV ?1HIV-positivemothers of babies born early diagnosis ;the fourth generation of HIV ?1antigen / antibodyELISA reagent for the detection of positive response ,but HIV ?1 antibodyconfirmation .
Auxiliary diagnosis ;monitoring disease progression and efficacy of antiviral therapy .But the existing monitoring disease progression and efficacy of antiviral therapy are expensive ,complex virus RNA determination method .
Chinese HIV/AIDS patients are mostly at the grassroots level, detection of 1viral loadto on 1000 yuan ( about 15 dollars) ,it is difficult to popularize in basic level ,for the treatment of diseases and control is very adverse .
High sensitivity p24 antigen detection method be used in developing viral load determination method of a choice .In recent years ,foreign to establish high sensitivity for the detection of p24 antigen inhas not stopped ,in order to improve the detection of serum p24 sensitivity to antigen,the immune complexes in serum after determination of dissociation ,developed by ICD p24 antigen determinationmethod .
23 years immune complex thermal dissociation via TSA signal amplification system using ELISA were detected, the detection of p24 antigen minimum detection by original 1 pg/mL down to.5 pg/mL,in HIV ?1HIV-positiveinfants born to mothers with early diagnosis and detection of RNA phase when ,with HIV nucleic acid detection with comparability ,it has important practical value .
For 24 years ,from the HIV virus found in the United States one of the University of Maryland Dr.Robert C.Gallo led by the laboratory of a high sensitivity for the detection of p24 antigen research has attracted wide attention ,p24 antigen detection becomes more sensitive than virus nucleic acid testing method .
25 years from University of Maryland in the United States, a long review also began to focus on p24 antigen detection,it can be used as a low-cost ,suitable for use in developing an alternative to the existing expensive RNA method for the determination of a choice .
At present, commercial p24 antigenstandard detection method is mainly based on the principle of sandwich ELISA .The method is used for anti p24 antigenantibody coated solid phase double antibody sandwich method ,is the most commonly used method of detecting antigens ,home is no commercial HIV p24 antigen detection reagentproduction .
To sum up ,with high sensitive analysis method for the diagnosis of AIDS will contribute to early diagnosis ,to avoid missed ,contribute to the treatment of AIDS drug efficacy evaluation ,prediction and monitoring of the disease process ,improve the detection sensitivity ,the shortening of the window period ,is the HIV diagnosis method and diagnosis reagent for the main direction of the development of .
Above the HIV epidemic survey and laboratory detection technology ,with HIV laboratory detection technology for continuous improvement ,particularly nucleic acid detection technology, HIV detection window period will be shortened gradually ,spread by blood transfusion will greatly reduce the possibility of HIV .
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8 WeberB ,Gurtler L ,Thorstensson R ,et al.Multicenter evaluation of a new automated fourth ?Generation human immunodeficiency virus screening assay with a sensitive antigen detection module and high J Clin Microbiol ,2 2,4(6):1938?1946.
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HIV ?1/HIV?2 ofWB J AIDS ,1994,7(6):623.11 GroopmanJE. Current advances in the diagnosis and treatment of AIDS: an Rev Infect Dis ,1999,12(5):98?911.12 ZhaoY ,Yu M ,Miller JW ,et al.
Quantification of human immunodeficiency virus type 1 proviralDNA by using TaqMan J Clin Microbiol 2,4,2(2):675?678.13 OU ,Swiggard WJ ,Jeyakumar D ,et al. A sensitive ,quantitative assay for human immunodeficiency virus type 1 integrat J Virol ,2 2,76(21):1942?195.
14 ToddJ ,Pachl C ,White R ,et al. Performance characteristics for the quantitation of plasma HIV RNA using branched DNA ?1signalamplification J Acquir Immune Defic Syndr Hum Retrovirol 5,1,2(2):35?44.
15LuoLi-min ,Huang Jiansheng ,Li Ming, application of quantitative detection of NASBA HCVRNA research Journal of Tropical Medicine ,2(1):642,2?66.16BaranyF. Genetic disease detection and DNA amplification using cloned thermostable Proc Natl Acad Sci U S A ,1991,88(1):189?193.
17Wei Min.HIV testing laboratory research progress and development trend Foreign Medical Sciences ( virology section) ,2 3,1(3):65?69.18 MylonakisE ,Lally M ,Lally M. Laboratory testing for infection with human immunodeficiency virus: established and novel Journal OF MEDICINE ,2,19(7):568?576.
19 GurtlerL ,Michl U ,Michl U. Reduction of the diagnostic window with a new combined p24 antigenand humar immunodeficiency virus antibody screening Journal of Virological Methods ( 1,1998,7538.
2 ):27?Weber B ,Rabenau H ,Rabenau H. Evaluation of a New Combined Antigen and Antibody Human Immunodeficiency virus screening assay ,VIDAS HIV DUO Journal of Clinical Microbiology 2,4,2(4):142?1426.
21 WeberB ,Thorstensson R ,Thorstensson R. Multicenter Evaluation of a new automated fourth ?Generation human immunodeficiency virus screening assay with a sensitive antigen detection module and high Journal of Clinical Microbiology ,22,4(6):1938?1946.
22 Xu Keyi,Wang Ying .The simultaneous detection of HIV antibody and p24 antigenrapid diagnosis reagent Chinese Journal of experimental and clinical virology ,2(4):3772,16379.23Shen XiaAIDS ? Chinese Journal of laboratory medicine ,2 3,26(5):327?328.
24 and Zhao Liqing,Li Lianxue. ELISA was detected in the urine HIV ?Ⅰantibodyanalysis China AIDS STD prevention and control ,2 3,9(5):276?277.25Feng Xiaoyingdrug addicts urine specimens of blood HIV ?ELISA Ⅰantibodytesting and comparative analysis STD AIDS ,2 3,9(3):132?133.
26 MeierT ,Henkes M ,Henkes M.Eviden CE for a diagnostic window in fourth generation assays for Journal of Clinical Virology ,2 1,23:113?116.27 Sutthent,Jeyakumar D. A sensitive ,quantitative assay for human immunodeficiency virus type 1 Journal of Virology 3,76,2(21):1942?195 aidsrapid ( gold standard ) detection of colloidal gold ( gold standard) of human immunodeficiency virus ( HIV 1/2)antibody detection kit is the application of colloidal gold immune chromatography technology to establish a sensitive ,specific ,simple one-step HIV 1/2 antibodyqualitative testing agent .
For qualitative detection of human serum or plasma samples of HIV 1and HIV 2specific antibodies,to assist in the diagnosis of HIV infection ,the positive results using immunoblotting confirmed .
The detection principle is using HIV recombinant antigen and Sheep anti mouse IgG were immobilized on a nitrocellulose membrane ,and coated with colloidal gold labeled recombinant antigens of the HIV and rat IgG glass fiber paper and other reagents .
Application of colloidal gold immunochromatographic technique ,using the principle of the double antigen sandwich assay of human serum and plasma samples of HIV 1/2 antibody.The detection process, blood samples were added to the kit for the sample adding hole ,first samples and glass fiber paper of colloidal gold labeled recombinant antigens of the HIV are mixed ,and then to the cellulose nitrate membrane chromatography .
If the sample contains HIV 1/2 antibodies,these antibodies ,and package is HIV recombinant antigen for colloidal gold combined ,such mixtures to nitrocellulose membrane chromatography ,can be fixed with recombinant HIV antigen detection line ( T line) capture and the formation of colloidal gold labeled antigen - antibody - antigen double antigen sandwich immunoassay complex ,therefore in the T line appears a red line ,for positive results .
If the subject is not HIV 1/2 antibodyin the blood ,is not in the test line ( T line) is formed on the red line ,negative results .In the kit on the control line ( C line) coated with Sheep anti mouse IgG ,under no circumstance will with rat IgG- colloidal gold combined in the quality control on the red line ,to prove that the kit is working .
Its characteristics are convenient ,rapid (1 min results ) ,accurate ,is the preferred method of screening for AIDS patients .Rapid detection of products in two is a blood test is a blood test strip saliva test strip general Abbott ,Palmer and saliva test is only a kind of the United States Awel saliva detection test paper production base in the mano Beijing biotechnology limited formal AIDS detection test, batch production can be the State Food and drug administration the query is also had AIDS test fakes such situations .
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2011-05-15
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