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[应用聚合酶链反应检测血清乙型肝炎病毒DNA]

[Detection of serum hepatitis B virus DNA using the polymerase chain reaction assay].

作者信息

Kaneko S, Unoura M, Kobayashi K

机构信息

1st Department of Internal Medicine, Kanazawa University.

出版信息

Rinsho Byori. 1990 Sep;38(9):1036-40.

PMID:2232264
Abstract

We compared the sensitivity of the polymerase chain reaction (PCR) assay to that of slot blot hybridization for detecting hepatitis B virus (HBV) DNA in the serum of a chimpanzee infected with HBV and 52 patients. Also, we utilized a rapid PCR procedure for the detection: Viral DNA was released from virions by incubating serum with NaOH. After a primary PCR amplification, the sample was reamplified using a second set of primer pairs (PCR/PCR). In the chimpanzee, HBV DNA was detected 3 weeks earlier than the appearance of hepatitis surface antigen (HBsAg) and persisted for two weeks with antibody to HBsAg. Of the 14 chronic hepatitis B patients positive for both HBsAg and HBV e antigen (eAg), 9 were positive for HBV DNA by slot blot hybridization and all 14 by PCR. Also, of 9 patients positive for HBsAg and antibody to eAg, 2 were positive for HBV DNA by slot blot hybridization and 8 by PCR. Three of the 11 patients who had lost HBsAg during follow up examination of chronic hepatitis B were positive for HBV DNA by PCR, whereas none of them was positive by slot blot hybridization. Six patients who had recovered from acute hepatitis B more than one year ago and 12 cases who had had vaccination of HBV were negative for HBV DNA by PCR. This technique should yield valuable information on the biology of HBV.

摘要

我们比较了聚合酶链反应(PCR)检测法与狭缝印迹杂交法对感染乙肝病毒(HBV)的黑猩猩血清及52例患者血清中HBV DNA的检测灵敏度。此外,我们采用了一种快速PCR检测程序:通过将血清与氢氧化钠孵育从病毒颗粒中释放病毒DNA。经过一次PCR扩增后,使用第二组引物对再次扩增样品(PCR/PCR)。在黑猩猩中,比乙肝表面抗原(HBsAg)出现早3周检测到HBV DNA,并且在出现HBsAg抗体后持续两周。在14例HBsAg和乙肝e抗原(eAg)均呈阳性的慢性乙型肝炎患者中,9例通过狭缝印迹杂交法检测HBV DNA呈阳性,而14例通过PCR检测均呈阳性。另外,在9例HBsAg和eAg抗体呈阳性的患者中,2例通过狭缝印迹杂交法检测HBV DNA呈阳性,8例通过PCR检测呈阳性。在慢性乙型肝炎随访检查中失去HBsAg的11例患者中,3例通过PCR检测HBV DNA呈阳性,而通过狭缝印迹杂交法检测均为阴性。6例一年多前从急性乙型肝炎康复的患者和12例接种过乙肝疫苗的患者通过PCR检测HBV DNA均为阴性。这项技术应该能提供有关乙肝病毒生物学的有价值信息。

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