论文部分内容阅读
在人巨细胞病毒（ＨＣＭＶ）－ＡＤ＿（１６９）株直接早期基因的ＥｃｏＲＩＪ片段内，自行设计二对引物，建立套式聚合酶链反应（ＰＣＲ）加限制酶分析检测ＨＣＭＶＤＮＡ，经实验证明有高度的特异性与敏感性，对其它疽疹类病毒及正常人基因组ＤＮＡ无交叉反应。一次性ＰＣＲ检测ＨＣＭＶ－ＡＤ１６９株基因组的最小检出量为１００ｆｇ，而套式ＰＣＲ可达１０ｆｇ。经ＰｓｔＩ酶切后，得到的片段与设计相符。对５８名孕早期孕妇外周静脉血进行ＨＣＭＶＤＮＡ、病毒分离、特异性ＩｇＭ及ＩｇＡ检测，结果阳性率分别８．６％、６．９％、１．７％及３。４％。２８例死胎组织中，发现一死胎肺组织ＨＣＭＶＤＮＡ阳性，与病毒分离相比，套式ＰＣＲ的特异性，敏感性及符合率分别达到１００％，９８．１％及９８．３％。提示：套式ＰＣＲ能提高诊断ＨＣＭＶ感染的特异性与敏感性，适合于孕早期ＮＣＭＶ感染的监测。
Two pairs of primers were designed in the EcoRIJ fragment of direct early gene of human cytomegalovirus (HCMV) -AD169 strain to establish nested polymerase chain reaction (PCR) and restriction enzyme analysis to detect HCMVDNA. Specificity and sensitivity, no other cross-reactivity to other rash virus and normal human genomic DNA. One-time PCR detection of HCMV-AD169 genome minimum detection of 100fg, and nested PCR up to 10fg. After digestion with PstI, the fragments obtained are in accordance with the design. The positive rate of HCMVDNA, virus isolation, specific IgM and IgA in 58 pregnant women with early pregnancy was 8.6%, 6.9%, 1.7% and 3.4% respectively. In 28 cases of stillbirth, HCMVDNA was found in the fetal lung tissue. Compared with the virus, the specificity, sensitivity and coincidence rate of the nested PCR were 100%, 98.1% and 98.3% respectively. Tip: Nested PCR can improve the diagnosis of HCMV infection specificity and sensitivity, suitable for early pregnancy NCMV infection monitoring.