Garcia F, Garcia F, Bernal M C, Leyva A, Piedrola G, Maroto M C
Microbiology Department, Faculty of Medicine, University of Granada, Spain.
J Clin Microbiol. 1995 Feb;33(2):413-5. doi: 10.1128/jcm.33.2.413-415.1995.
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.
我们评估了一种新的酶免疫分析技术，用于检测基于聚合酶链反应（PCR）扩增的产物，该技术可能适用于乙型肝炎病毒（HBV）DNA的常规检测。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。我们将这种新的检测方法（PCR-DNA酶免疫分析[DEIA]）与未经事先PCR扩增的斑点印迹杂交以及另外两种检测PCR产物的方法进行了比较：溴化乙锭染色琼脂糖凝胶电泳（PCR-EB）和斑点印迹（PCR-斑点印迹）。对于乙型肝炎抗原阳性基础样本，斑点印迹法检测到HBV DNA的比例为70.4%，PCR-EB法为74.1%，PCR-DEIA法和PCR-斑点印迹法均为100%；对于抗乙型肝炎e抗原基础样本，斑点印迹法和PCR-EB法检测到HBV DNA的比例为10.5%，PCR-DEIA法和PCR-斑点印迹法为42.1%。卡方检验显示斑点印迹与PCR-EB之间以及PCR-DEIA与PCR斑点印迹之间存在强关联。以PCR-斑点印迹为参照，斑点印迹的灵敏度为56.9%，特异性为100%，PCR-EB的灵敏度为55.0%，特异性为100%，PCR-DEIA的灵敏度为95.4%，特异性为97%。我们得出结论，DEIA方法的技术优势及其高灵敏度和特异性可能有助于临床微生物实验室在HBV DNA常规检测中应用PCR技术。
