本文選題：乙肝病毒　切入點(diǎn)：反向斑點(diǎn)雜交　出處：《中國(guó)現(xiàn)代醫(yī)學(xué)雜志》2015年03期 　論文類型：期刊論文

【摘要】：目的基于多重PCR和反向斑點(diǎn)雜交原理,建立快速檢測(cè)乙肝病毒(HBV)基因型和耐藥性的膜芯片方法,并對(duì)其臨床應(yīng)用價(jià)值進(jìn)行探討。方法根據(jù)HBV基因序列設(shè)計(jì)探針并制作膜芯片,用PCR-反向斑點(diǎn)雜交檢測(cè)3種HBV標(biāo)準(zhǔn)菌株和38例HBV臨床分離株的基因型和耐藥性,檢測(cè)結(jié)果與測(cè)序結(jié)果進(jìn)行比較,分析方法的準(zhǔn)確性和最低檢出限。對(duì)深圳地區(qū)187例HBV陽(yáng)性標(biāo)本進(jìn)行基因分型和耐藥性檢測(cè),了解其臨床應(yīng)用價(jià)值。結(jié)果 38例HBV臨床分離株和3株HBV標(biāo)準(zhǔn)株鑒定結(jié)果與測(cè)序法完全一致,3株非HBV病毒和11株非HBV標(biāo)準(zhǔn)菌株無(wú)陽(yáng)性顯色,該方法準(zhǔn)確性達(dá)到100%,最低檢出限為103 copies/ml。深圳地區(qū)187例HBV臨床分離株以B型為主,占57.8%,C型占37.4%,未測(cè)出D型。187例HBV臨床分離株的耐藥突變率為26.2%,以rt L180M+rt M204V/I為主,占67.3%,其次為rt M204I,占16.3%。臨床HBV耐藥株以C型為主,占65.3%,B型占34.7%。結(jié)論用膜芯片檢測(cè)HBV基因型與耐藥性具有簡(jiǎn)便、快速、準(zhǔn)確性及特異性高的優(yōu)點(diǎn),評(píng)價(jià)指標(biāo)符合臨床實(shí)驗(yàn)要求,可在臨床推廣使用。
[Abstract]:Objective based on the principle of multiplex PCR and reverse dot blot hybridization, a method for rapid detection of HBV genotypes and drug resistance was established, and its clinical application value was discussed. Methods the probe was designed and the microarray was made according to the sequence of HBV gene. The genotypes and drug resistance of 3 HBV standard strains and 38 clinical isolates of HBV were detected by PCR- reverse dot blot. The results were compared with those of sequencing. The accuracy and minimum detection limit of the method were analyzed. The genotyping and drug resistance of 187 HBV positive samples from Shenzhen area were detected. Results the identification results of 38 HBV clinical isolates and 3 HBV standard strains were completely consistent with the sequencing method. No positive staining was found in 3 non-#en2# virus strains and 11 non-#en3# standard strains. The accuracy of this method was 100. The lowest detection limit was 103copias / ml. In Shenzhen area, 187 clinical isolates of HBV were mainly type B, accounting for 57.8% and C type, and the mutation rate of drug resistance in 187 clinical isolates of type D was 26.20.The main type was RT L180M rt M204V / I. The clinical HBV resistant strains were mainly type C, accounting for 65.3% of B type and 34.70.Conclusion Detection of HBV genotypes and drug resistance by membrane microarray is simple, rapid, accurate and specific, and the evaluation indexes meet the requirements of clinical experiments. It can be widely used in clinic.
【作者單位】： 深圳市人民醫(yī)院;
【基金】：深圳市科技工貿(mào)和信息化委員會(huì)基金資助項(xiàng)目(No:201103360)
【分類號(hào)】：R512.62

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