EVALUATION OF ENZYME-IMMUNOASSAY FOR HEPATITIS-B VIRUS-DNA BASED ON ANTI-DOUBLE-STRANDED DNA
EVALUATION OF ENZYME-IMMUNOASSAY FOR HEPATITIS-B VIRUS-DNA BASED ON ANTI-DOUBLE-STRANDED DNA
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DOI：
10.1128/jcm.33.2.413-415.1995
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发表时间：
1995-02-01
影响因子：
9.4
通讯作者：
MAROTO, MC
中科院分区：
文献类型：
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作者：
GARCIA, F;GARCIA, F;MAROTO, MC
我们评价了一种新的酶免疫分析技术，用于检测PCR扩增产物，该技术可能适用于常规检测B型肝炎病毒（HBV）DNA。研究了208份血清样本：73份为基础样本，135份为慢性肝炎患者的连续血清样本，其中一些患者正在接受α干扰素治疗。我们比较了新的检测方法（PCR-DNA酶免疫测定法[DEIA]）与斑点杂交进行事先没有PCR扩增，并与其他两种方法检测PCR产物：琼脂糖凝胶电泳与溴化乙锭染色（PCR-EB）和斑点杂交（PCR-dot blot）。对乙肝抗原阳性的基础标本，斑点杂交法、PCR-EB法、PCR-DEIA法和PCR-dotblot法的检出率分别为70.4%、74.1%、100%，对抗乙肝B e抗原的基础标本，斑点杂交法和PCR-EB法的检出率分别为10.5%、42.1%。卡方检验显示斑点杂交与PCR-EB之间以及PCR-DEIA与PCR斑点杂交之间存在强相关性。以PCR-dot blot为参照，dot blot的敏感性为56.9%，特异性为100%，PCR-EB的敏感性为55.0%，特异性为100%，PCR-DEIA的敏感性为95.4%，特异性为97%。我们的结论是，DEIA方法的技术优势和它的高灵敏度和特异性，可能有助于在临床微生物实验室的常规检测HBV DNA的PCR的使用。
We have evaluated a new enzyme immunoassay technology to detect the products of PCR-based amplification that may be applicable to routine testing of hepatitis B virus (HBV) DNA. Two hundred eight serum samples were studied: 73 were basal samples and 135 were sequential serum samples from patients with chronic hepatitis, some of whom were being treated with alpha interferon. We compared the new detection method (PCR-DNA enzyme immunoassay [DEIA]) with dot blot hybridization performed without prior PCR amplification and with two other methods for detection of PCR products: agarose gel electrophoresis with ethidium bromide staining (PCR-EB) and dot blot (PCR-dot blot). For hepatitis B-antigen-positive basal samples, HBV DNA was detected in 70.4% by dot blot, 74.1% by PCR-EB, and 100% by PCR-DEIA and PCR-dot blot; for anti-hepatitis B e-antigen basal samples, HBV DNA was found in 10.5% by dot blot and PCR-EB and in 42.1% by PCR-DEIA and PCR-dot blot. Chi-square tests showed a strong association between dot blot and PCR-EB and between PCR-DEIA and PCR dot blot. Using PCR-dot blot as the reference, dot blot shows a 56.9% sensitivity and a 100% specificity, PCR-EB shows a 55.0% sensitivity and a 100% specificity, and PCR-DEIA shows a 95.4% sensitivity and a 97% specificity. We conclude that the technical advantages of the DEIA method and its high sensitivity and specificity may facilitate the use of PCR in routine testing for HBV DNA in clinical microbiology laboratories.