本文選題：1型艾滋病病毒 + 核酸定性檢測(cè)　； 參考：《中國(guó)疾病預(yù)防控制中心》2017年碩士論文

【摘要】：目前常用的核酸定性檢測(cè)技術(shù)包括:實(shí)驗(yàn)室自建的In-house方法、Abbott M2000HIV-1 RNA/DNA以及AptimaHIV-1RNA核酸定性檢測(cè)。常用的核酸定量檢測(cè)技術(shù)包括羅氏診斷的Cobas Amplipre/Cobas TaqMan HTV-1 Testv 2.0、雅培公司的RealTime HIV-1、梅里埃公司的 Nuclisens EasyQ HIV-1 v2.0 和西門子公司的 Versant HIV-1 RNA 3.0 assay(b-DNA)。隨著分子生物學(xué)技術(shù)的發(fā)展,更靈敏的核酸檢測(cè)方法應(yīng)運(yùn)而生,包括數(shù)字PCR技術(shù)等。本研究采用Nuclisens EasyQ HIV-1核酸定量檢測(cè)、PCR 熒光探針法和數(shù)字 PCR(digita1 polymerase chain reaction,dPCR)對(duì) HIV診斷及治療后進(jìn)行了系統(tǒng)研究,主要包括以下三個(gè)部分:第一部分核酸定量檢測(cè)采用梅里埃公司的Nuclisens EasyQ HIV-1 v2.0,直接檢測(cè)血漿中HIV的RNA量。全面地分析核酸定性以及定量檢測(cè)的結(jié)果,深入地探討在實(shí)驗(yàn)室中,核酸定量檢測(cè)用于診斷HIV的可行性以及可操作性。第二部分采用PCR-熒光探針法檢測(cè)HIV患者治療后PBMCs內(nèi)HIV-1前病毒DNA的水平,探索在血漿HIV-1 RNA低于檢測(cè)下限時(shí),血細(xì)胞HIV-1前病毒作為判斷抗病毒治療療效以及病情進(jìn)展預(yù)測(cè)指標(biāo)的價(jià)值,為進(jìn)一步探索清除病毒的新治療措施奠定基礎(chǔ)。第三部分采用ddPCR、qPCR對(duì)不同月齡的HIV感染新生兒進(jìn)行檢測(cè),比較兩種方法的靈敏度和特異度;并評(píng)價(jià)ddPCR、qPCR兩種檢測(cè)方法的一致性,為制定我國(guó)的HIV暴露嬰兒的實(shí)驗(yàn)室診斷檢測(cè)策略做參考依據(jù)。第一部分核酸定量檢測(cè)應(yīng)用于HIV-1感染診斷的研究研究背景艾滋病(Acquired immune deficiency syndrome,AIDS)的病原體是人類免疫缺陷病毒(Human Immunodeficiency Virus,HIV),2017 年 2 月,全國(guó)(不含港澳臺(tái))共報(bào)告法定傳染病485649例,死亡1409例,其中AIDS死亡人數(shù)1097例(77.9%)。將HIV及時(shí)地發(fā)現(xiàn)并且及早期地診斷出對(duì)艾滋感染者的治療非常緊要,重要的是它能夠有效地降低患者的死亡率、并且能夠?qū)⑵渖娴馁|(zhì)量提高。艾滋病目前常用的檢測(cè)手段主要有抗體檢測(cè)和病毒檢測(cè),病毒檢測(cè)主要包括p24抗原檢測(cè)、細(xì)胞培養(yǎng)(病毒分離)以及核酸檢測(cè)�？贵w檢測(cè)通常是在感染后3-8周才能檢測(cè)出來,而核酸檢測(cè)能夠在感染后2周內(nèi)檢測(cè)出來,從而縮短窗口期。核酸檢測(cè)的好處有兩個(gè)即敏感性以及特異性較高,因此能夠作為艾滋早期診斷的重要手段。根據(jù)《全國(guó)艾滋病檢測(cè)技術(shù)規(guī)范》(2015年)修訂版中的規(guī)定:檢測(cè)策略補(bǔ)充試驗(yàn)包括抗體確證以及核酸試驗(yàn),將核酸檢測(cè)提到了艾滋病檢測(cè)診斷的關(guān)鍵地位。定量檢測(cè)能夠?qū)⑼庵苎械腍IV病毒含量測(cè)出,比定性檢測(cè)更加便捷和敏感。本研究中核酸定性檢測(cè)采用中國(guó)疾病預(yù)防控制中心性病艾滋病中心參比實(shí)驗(yàn)室自建的In-house方法,應(yīng)用人類所有細(xì)胞均含有的β-actin基因作內(nèi)參,當(dāng)先擴(kuò)增出β-actin片段后再根據(jù)巢式的方法用pol區(qū)、gag區(qū)、env區(qū)相對(duì)應(yīng)的引物來擴(kuò)增HIV特異性的基因片段,最終結(jié)果根據(jù)擴(kuò)增產(chǎn)物來判定。核酸定量檢測(cè)則采用梅里埃公司的Nuclisens EasyQ HIV-1 v2.0,直接檢測(cè)血漿中HIV的RNA量。全面地分析核酸定性以及定量檢測(cè)的結(jié)果,深入地探討在實(shí)驗(yàn)室中,核酸定量檢測(cè)用于診斷HIV的可行性以及可操作性。目的探討核酸定量檢測(cè)在1型艾滋病病毒(HIV-1)實(shí)驗(yàn)室感染診斷中的應(yīng)用。對(duì)象和方法選取云南省德宏州216例樣本,包括136例HIV-1陽性母親所生活產(chǎn)嬰兒和80例HIV-1陽性成人,分別用HIV-1核酸定性和定量方法進(jìn)行檢測(cè),綜合對(duì)比分析2種方法的檢測(cè)結(jié)果。結(jié)果1.216例樣本的核酸定性和定量檢測(cè)試驗(yàn)一致性極好,53例樣本(50例成人樣本和3例嬰兒樣本)核酸陽性,血漿病毒載量5000CPs/mL,隨訪50例成人樣本的蛋白印跡試驗(yàn)(WB)確證均為HIV-1抗體陽性;2.163例陰性樣本(30例成人樣本和133例嬰兒樣本)核酸陰性,血漿病毒載量均低于檢測(cè)下限。結(jié)論當(dāng)血漿病毒載量5000CPs/mL,核酸定量檢測(cè)試驗(yàn)對(duì)于HIV-1感染的陽性樣本是一種有效的實(shí)驗(yàn)室診斷方法。第二部分PCR-熒光探針法在成人HIV/AIDS接受長(zhǎng)期HAART治療后的應(yīng)用研究研究背景人體感染HIV后可整合在宿主基因上,從而使病毒潛伏在淋巴細(xì)胞,腦細(xì)胞以及消化道淋巴組織等組織器官中,形成了病毒儲(chǔ)存庫,高效抗逆轉(zhuǎn)錄病毒治療不能徹底清除人體內(nèi)的病毒。病毒潛伏或潛伏病毒即病毒在被感染的細(xì)胞中暫不復(fù)制,可逃避宿主的免疫清除,但在一定的條件下可復(fù)制病毒。HAART能夠很明顯地降低艾滋病的病死率,也能夠抑制病毒的有效復(fù)制,同時(shí)不僅將HIV/AIDS血漿中的病毒載量降低至目前現(xiàn)存的檢測(cè)方法的最低檢測(cè)水平以下,而且還能升高CD4+T細(xì)胞數(shù)量,很好地重新構(gòu)建人體的免疫功能。但是,至今為止尚不能徹底治愈HIV,可能是因?yàn)槿梭w內(nèi)會(huì)持續(xù)地存在病毒儲(chǔ)存庫。本研究采用PCR-熒光探針法檢測(cè)HIV患者治療后PBMCs內(nèi)HIV-1前病毒DNA的水平,擬探索在血漿HIV-1 RNA低于檢測(cè)下限時(shí),將PBMCs細(xì)胞內(nèi)HIV-1前病毒水平作為判斷抗病毒治療療效以及病情進(jìn)展預(yù)測(cè)指標(biāo)的價(jià)值,為進(jìn)一步探索清除病毒的新治療措施奠定基礎(chǔ)。目的探討PCR-熒光探針法檢測(cè)成人HIV/AIDS接受高效抗逆轉(zhuǎn)錄病毒治療(HAART)后HIV-1病毒儲(chǔ)存庫的改變情況及病毒儲(chǔ)存庫與治療效果的關(guān)系。對(duì)象和方法對(duì)91例規(guī)范接受HAART治療5-10年的HIV-1患者采集血標(biāo)本進(jìn)行橫斷面研究;通過分支DNA(bDNA)法檢測(cè)血漿HIV-1RNA病毒載量;實(shí)驗(yàn)室自建的In-house巢式PCR、PCR-熒光探針法檢測(cè)外周血HIV-IDNA水平;流式細(xì)胞術(shù)檢測(cè)外周血CD4+T細(xì)胞數(shù)量;對(duì)數(shù)據(jù)進(jìn)行統(tǒng)計(jì)學(xué)處理。結(jié)果1.91例接受5-10年HAART治療的HIV-1患者外周血HIV-1RNA均低于檢測(cè)下限(50拷貝/ml);2.細(xì)胞中HIV-1前病毒DNA的平均含量為1.77±0.76(log10拷貝/106個(gè)細(xì)胞);3.實(shí)驗(yàn)室自建的In-house巢式PCR法檢測(cè)細(xì)胞中HIV-1 DNA陰性者比例為39/91,不確定者比例為52/91;4.CD4+T細(xì)胞平均數(shù)量為479.56±188.11個(gè)/μl。結(jié)論P(yáng)CR-熒光探針法檢測(cè)經(jīng)過長(zhǎng)期HAART治療(5-10年)成人患者的病毒儲(chǔ)存庫更加靈敏,儲(chǔ)存庫的檢測(cè)對(duì)于抗病毒治療療效的判斷以及病情進(jìn)展的預(yù)測(cè)都有重要的意義。第三部分?jǐn)?shù)字PCR和熒光定量PCR用于HIV嬰兒早期診斷的研究研究背景HIV-l DNA可以整合到宿主DNA中,即為前病毒。而細(xì)胞內(nèi)總的HIVDNA是測(cè)量?jī)?chǔ)存庫大小的指標(biāo)。HIVDNA對(duì)于HIV的診斷、療效監(jiān)測(cè)具有很重要的意義。目前HIV DNA定量檢測(cè)主要采用實(shí)時(shí)熒光定量PCR(real—time quantitation PCR,qPCR),其依賴的定量基礎(chǔ)循環(huán)閾值(cycle threshold,Ct)受擴(kuò)增效率的影響很大,故qPCR只能相對(duì)定量。微滴式數(shù)字PCR(droplet digital polymerase chain reaction,ddPCR)是一種核酸檢測(cè)和絕對(duì)定量的新方法,將樣品通過微滴發(fā)生器進(jìn)行微滴化處理,形成數(shù)以萬計(jì)的油包水小液滴,每個(gè)液滴中含有不超過一個(gè)DNA分子,每個(gè)DNA分子均作為單獨(dú)的PCR反應(yīng)體系,經(jīng)PCR擴(kuò)增后與熒光標(biāo)記探針雜交,用微滴檢測(cè)器對(duì)每個(gè)反應(yīng)進(jìn)行檢測(cè),實(shí)現(xiàn)對(duì)單分子核酸的絕對(duì)定量。ddPCR技術(shù)無需建立標(biāo)準(zhǔn)曲線,且具有較高的靈敏度,但因其價(jià)格較高,所以國(guó)內(nèi)還沒有其在HIV定量檢測(cè)方面的研究。本研究擬采用ddPCR、qPCR對(duì)不同月齡的HIV感染新生兒進(jìn)行檢測(cè),比較兩種方法的靈敏度和特異度,并評(píng)價(jià)ddPCR、qPCR兩種檢測(cè)方法的一致性;為制定我國(guó)的HIV暴露嬰兒的實(shí)驗(yàn)室診斷檢測(cè)策略做參考依據(jù)。目的比較ddPCR、qPCR對(duì)不同月齡的HIV感染新生兒檢測(cè)的靈敏度與特異度,并評(píng)價(jià)ddPCR、qPCR兩種檢測(cè)方法的一致性。對(duì)象和方法選取2014-2016年云南、四川、新疆、重慶4省的HIV-1陽性母親所生活產(chǎn)嬰兒116例不同時(shí)間點(diǎn)干血斑(DBS)樣本,根據(jù)《全國(guó)艾滋病檢測(cè)技術(shù)規(guī)范》2015年修訂版,本研究的金標(biāo)準(zhǔn)為連續(xù)兩次核酸定性檢測(cè)為陽性結(jié)果即判定為陽性。即116例嬰兒干血斑樣本中,有54例陽性樣本,62例陰性樣本。結(jié)果1.116例嬰兒樣本,qPCR的靈敏度為63.0%,特異度為91.9%;ddPCR的靈敏度為59.2%,特異度為75.8%;qPCR與ddPCR檢測(cè)一致率為73.3%,Kappa=0.391;2.對(duì)于產(chǎn)后24h內(nèi)的樣本而言,qPCR的靈敏度為36.3%,特異度為80.0%;ddPCR的靈敏度為27.2%,特異度為73.3%;qPCR與ddPCR檢測(cè)一致率為53.8%,Kappa=-0.173;3.對(duì)于產(chǎn)后4w內(nèi)的樣本,qPCR的靈敏度為50.0%,特異度為100.0%;ddPCR的靈敏度為57.1%,特異度為92.9%。qPCR與ddPCR檢測(cè)一致率為78.6%,Kappa=0.478;4.對(duì)于產(chǎn)后6W樣本,qPCR的靈敏度為78.6%,特異度為100%;ddPCR的靈敏度為78.6%,特異度為77.8%。qPCR與ddPCR檢測(cè)一致率為87.5%,Kappa=0.745;5.對(duì)于產(chǎn)后3m樣本qPCR的靈敏度為80.0%,特異度為86.7%;ddPCR的靈敏度為66.7%,特異度為60.0%。qPCR與ddPCR檢測(cè)一致率為66.7%,Kappa=0.336。結(jié)論ddPCR與qPCR兩種檢測(cè)方法的靈敏度與特異度均不高,且兩種檢測(cè)方法一致性較差,暫不能作為嬰兒HIV早期診斷檢測(cè)的參考依據(jù)。
[Abstract]:Currently, the commonly used nucleic acid detection techniques include the In-house method built by the laboratory, the Abbott M2000HIV-1 RNA/DNA and the qualitative detection of AptimaHIV-1RNA nucleic acid. The commonly used nucleic acid quantitative detection techniques include Roche's Cobas Amplipre/Cobas TaqMan HTV-1 Testv 2, the RealTime HIV-1 of the Abbott Company, and the mill of the company. EasyQ HIV-1 v2.0 and SIEMENS's Versant HIV-1 RNA 3 assay (b-DNA). With the development of molecular biology technology, more sensitive nucleic acid detection methods have come into being, including digital PCR technology, etc. This study uses Nuclisens EasyQ HIV-1 nucleic acid quantitative detection. On, dPCR) carried out a systematic study on the diagnosis and treatment of HIV, including the following three parts: the first part of the quantitative detection of nucleic acid, using the Nuclisens EasyQ HIV-1 v2.0 of the company, to determine the RNA amount of HIV in the plasma directly. The results of nucleic acid determination and quantitative detection are analyzed comprehensively. The nucleic acid quantification in the laboratory is deeply discussed. The feasibility and maneuverability of detecting HIV were detected. The second part used PCR- fluorescence probe to detect the level of HIV-1 virus DNA in PBMCs after HIV treatment, and explored the value of the pre HIV-1 virus of blood cells as the value of judging the curative effect of antiviral treatment and the predictor of the progression of the disease when the plasma HIV-1 RNA was lower than the detection limit. The third part uses ddPCR and qPCR to detect the HIV infected newborns with different months of age, compare the sensitivity and specificity of the two methods, and evaluate the consistency of the two methods of ddPCR and qPCR, and make reference to the formulation of laboratory diagnostic testing strategies for HIV exposed infants in China. The first part of the quantitative detection of nucleic acid is applied to the diagnosis of HIV-1 infection. The pathogen of Acquired immune deficiency syndrome (AIDS) is the human immunodeficiency virus (Human Immunodeficiency Virus, HIV). In February 2017, 485649 cases of statutory infectious diseases were reported in China (excluding Hong Kong, Macao and Taiwan), and 1409 cases died, of which A The number of IDS deaths is 1097 (77.9%). The timely detection and early diagnosis of HIV and the early diagnosis of AIDS infected people are very important. It is important that it can effectively reduce the death rate of the patient and improve the quality of its survival. The current detection hand of AIDS is mainly antibody detection and virus detection, virus detection. It mainly includes p24 antigen detection, cell culture (virus isolation) and nucleic acid detection. Antibody detection is usually detected at 3-8 weeks after infection, and nucleic acid detection can be detected within 2 weeks after infection, thus reducing the window period. The advantages of nucleic acid detection are two sensitivity and high specificity, so it can be used as early AIDS. An important means of phase diagnosis. According to the provisions of the National AIDS test specification (2015) revised edition: the test strategy supplementary test includes antibody confirmation and nucleic acid test, which refers to the key status of the detection and diagnosis of AIDS. Quantitative detection can detect the content of HIV virus in peripheral blood more than qualitative detection. It is convenient and sensitive. In this study, the nucleic acid qualitative detection used the In-house method built by the reference laboratory of the center for STD AIDS in the center for disease prevention and control of China. The beta -actin gene contained in all human cells was used as the internal reference, the beta -actin fragment was amplified first and then the corresponding primers in the pol region, gag region, and env region were used by the nested method. To amplify the specific gene fragment of HIV, the final result is determined according to the amplified product. The nucleic acid quantitative detection uses the Nuclisens EasyQ HIV-1 v2.0 of mrer company to directly detect the RNA quantity of HIV in the plasma. The results of nucleic acid qualitative and quantitative detection are analyzed comprehensively. The quantitative detection of nucleic acid in the laboratory is used to diagnose the nucleic acid in the laboratory. HIV's feasibility and maneuverability. Objective to explore the application of nucleic acid quantitative detection in the diagnosis of laboratory infection of type 1 AIDS virus (HIV-1). Objects and methods selected 216 samples from Dehong, Yunnan Province, including 136 cases of HIV-1 positive mothers living and 80 cases of HIV-1 positive adults, using HIV-1 nucleic acid qualitative and quantitative methods respectively. Results of the test, the results of the 2 methods were compared and analyzed. Results the consistency of the nucleic acid qualitative and quantitative test of 1.216 samples was excellent, 53 samples (50 adult samples and 3 infant samples) were positive for nucleic acid and plasma viral load 5000CPs/mL. The egg white blot test (WB) of 50 adult samples were confirmed to be HIV-1 antibody positive; 2.163 The negative samples (30 adult samples and 133 infant samples) were negative and the plasma viral load was lower than the lower detection limit. Conclusion when the plasma viral load is 5000CPs/mL, the nucleic acid quantitative detection test is an effective laboratory diagnosis method for the positive samples of HIV-1 infection. The second part of the PCR- fluorescence probe is accepted in adult HIV/AIDS. The application of HAART after treatment study background human infection HIV can be integrated on the host gene, which makes the virus latent in the tissues and organs such as lymphocytes, brain cells and digestive lymphatic tissues, and forms a virus store. High performance antiretroviral therapy can not clear the virus in the human body. Virus latent or latent disease Virus is not replicated in infected cells and escaping from the immune clearance of the host, but replicating virus.HAART under certain conditions can significantly reduce the mortality of AIDS and inhibit the effective replication of the virus, and not only reduce the viral load in the HIV/AIDS plasma to the most existing testing methods. Under low level of detection, the number of CD4+T cells can be increased and the immune function of the human body is rebuilt well. However, it is not possible to completely cure HIV so far. It may be because of the persistent presence of virus storage in the human body. This study used PCR- fluorescence probe to detect the level of HIV-1 pre virus DNA in PBMCs after the treatment of HIV patients. To explore the value of the level of HIV-1 pre virus in PBMCs cells as an indicator of the therapeutic effect of antiviral therapy and the predictor of progression of the disease in PBMCs cells when the plasma RNA is lower than the detection limit, and to explore the basis for further exploration of the new treatment measures for eliminating the virus. Objective to investigate the detection of adult HIV/AIDS by PCR- fluorescence probe for the acceptance of high performance antiretroviral drugs. The changes in the HIV-1 virus repository after treatment (HAART) and the relationship between the virus repository and the therapeutic effect. Objects and methods were used to conduct a cross-sectional study of the blood samples collected from 91 HIV-1 patients receiving HAART therapy for 5-10 years; the HIV-1RNA virus load in plasma was detected by the branch DNA (bDNA) method; the In-house nested PCR, PCR- fluore was built in the laboratory. The HIV-IDNA level of peripheral blood was detected by light probe, and the number of CD4+T cells in peripheral blood was detected by flow cytometry. The data were processed statistically. Results the peripheral blood HIV-1RNA of 1.91 cases of HIV-1 patients receiving 5-10 years of HAART treatment were lower than the lower detection limit (50 copies /ml), and the average content of HIV-1 pre virus DNA in 2. cells was 1.77 + 0.76 (log10 copy /106). 3. In-house nested PCR method was built in the laboratory to detect the proportion of HIV-1 DNA negative in the cells was 39/91, the ratio of the uncertain person was 52/91; the average number of 4.CD4+T cells was 479.56 + 188.11 / mu L. conclusion PCR- fluorescence probe was more sensitive and the storage library was detected after long-term HAART treatment (5-10 years). The judgment of the efficacy of antiviral therapy and the prediction of the progress of the disease are of great significance. Third parts of digital PCR and fluorescence quantitative PCR are used to study the early diagnosis of HIV infants. HIV-l DNA can be integrated into the host DNA, that is, the pre virus, and the total HIVDNA in the cell is a measure of the size of the repository.HIVDNA for HIV At present, the quantitative detection of HIV DNA mainly uses real-time quantitative PCR (real - time quantitation PCR, qPCR), and its dependent quantitative basic cycle threshold (cycle threshold, Ct) is greatly influenced by the amplification efficiency. Therefore, qPCR can only be relatively quantitative. Chain reaction, ddPCR) is a new method of nucleic acid detection and absolute quantification. The samples are microdripped by microdrop generator to form tens of thousands of small liquid droplets of oil water. Each droplet contains no more than one DNA molecule. Each DNA molecule is used as a separate PCR reaction system, and after PCR amplification with a fluorescent labeling probe Each reaction was detected by a micro drop detector. The absolute quantitative.DdPCR technology for single molecule nucleic acid was not required to establish a standard curve and had high sensitivity. But because of its high price, there was no study on quantitative detection of HIV in China. This study was intended to use ddPCR and qPCR for HIV infected newborns with different months of age. The sensitivity and specificity of the two methods were compared, and the consistency of the two methods of ddPCR and qPCR were evaluated and the reference basis for the laboratory diagnosis of HIV exposed infants in China. The purpose was to compare the sensitivity and specificity of ddPCR, qPCR to the detection of HIV infected newborns with different months of age, and to evaluate the two species of ddPCR and qPCR. Conformance of detection methods. Objects and methods selected 116 cases of HIV-1 positive mothers living in 4 provinces of Yunnan, Sichuan, Xinjiang, Chongqing and 116 cases of dry blood spots (DBS) at different time points. According to the National AIDS test specification >2015 revised edition, the gold standard of this study was the positive result of two consecutive tests of nucleic acid. There were 54 positive samples and 62 negative samples in the 116 samples of dry blood spots in 116 infants. The sensitivity of 1.116 infants was 63%, the specificity was 91.9%, the sensitivity of ddPCR was 59.2%, the specificity was 75.8%, and the coincidence rate of qPCR and ddPCR was 73.3%, Kappa=0.391; 2. for postpartum 24h samples, qPCR spirit. The sensitivity was 36.3%, the specificity was 80%, the sensitivity of ddPCR was 27.2%, the specificity was 73.3%, and the consistency between qPCR and ddPCR was 53.8%, Kappa=-0.173; 3. for postpartum 4W samples, the sensitivity of qPCR was 50%, the specificity was 100%; the sensitivity of ddPCR was 57.1%, and the specificity was 78.6%, Kappa=0.478; 4. for 92.9%.qPCR and ddPCR. After postpartum 6W samples, the sensitivity of qPCR was 78.6%, the specificity was 100%, the sensitivity of ddPCR was 78.6%, the coincidence rate of 77.8%.qPCR and ddPCR was 87.5%, Kappa=0.745; 5. for postpartum 3M samples, the sensitivity was 80%, the specificity was 86.7%, the sensitivity of ddPCR was 66.7%, the specificity of 60.0%.qPCR and ddPCR was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa was 66.7%, Kappa =0.336. conclusion the sensitivity and specificity of the two detection methods of ddPCR and qPCR are not high, and the consistency of the two detection methods is poor, which can not be used as the reference basis for the early diagnosis and detection of infant HIV.

【學(xué)位授予單位】：中國(guó)疾病預(yù)防控制中心
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R512.91

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