Hepatitis B virions isolated with antibodies to the pre-S1 domain reveal occult viremia by PCR in Alaska Native HBV carriers who have seroconverted.
Hepatitis B virions isolated with antibodies to the pre-S1 domain reveal occult viremia by PCR in Alaska Native HBV carriers who have seroconverted.
复制标题
用前 S1 结构域抗体分离的乙型肝炎病毒粒子通过 PCR 揭示了血清转化的阿拉斯加本地 HBV 携带者的隐匿性病毒血症。
DOI：
10.1046/j.1537-2995.2000.40080910.x
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发表时间：
2000
期刊：
影响因子：
2.9
通讯作者：
Vyas,GN
中科院分区：
文献类型：
--
作者：
Gandhi,MJ;Yang,GG;McMahon,BJ;Vyas,GN
背景：HBsAg出现之前或HBsAg消失之后发生的隐匿性病毒血症可通过基因扩增技术检测，其效率取决于核酸制备。研究设计和方法：为了从病毒血症血浆中分离HBV DNA，评价了用生物素化的前S1抗体偶联链霉亲和素包被的磁珠对完整HBV的免疫亲和捕获（IAC）。将IAC与硅胶法（Qiagen [QSG]）及其两种改进方法进行比较，其中将样品与裂解缓冲液在60 ℃下加热10分钟（QSG-60）或与蛋白酶-K在58 ℃下加热60分钟（QSG-PK）。通过半巢式PCR扩增HBV基因序列检测每份HBV DNA样本。共检测了36份编码血清样本，包括3份HBsAg阳性对照和33份血清转化（产生HBsAg抗体[抗-HBs]）的既往慢性HBV携带者。结果：在33例血清转换的前慢性HBV携带者中，IAC在17例样本中发现HBV DNA，而QSG-PK仅在11例样本中发现HBV DNA（p = 0.031），QSG-60仅在10例样本中发现HBV DNA（p = 0.016），QSG仅在9例样本中发现HBV DNA（p = 0.0078）。然而，17份样本在1：10稀释度下未扩增HBV DNA;因此，认为其具有低水平病毒血症。IAC显示HBV DNA早于或早于PHM 907、911和922面板中的其他方法。结论：IAC显然是HBsAg前窗口期患者HBV DNA扩增的最佳样本制备方法，也是检测血清转化（产生抗-HBs）的前慢性HBV携带者中持续低水平病毒血症的最佳方法。
BACKGROUND:Occult viremia occurring before the appearance of HBsAg or after the disappearance of HBsAg is detectable by gene amplification technologies whose efficiency depends on nucleic acid preparation.STUDY DESIGN AND METHODS:To isolate HBV DNA from viremic plasma, immunoaffinity capture (IAC) of intact HBV with biotinylated pre‐S1 antibodies coupled to streptavidin‐coated magnetic beads was evaluated. IAC was compared with a silica‐gel method (Qiagen [QSG]) and its two modifications wherein the samples were heated with lysis buffer at 60oC for 10 minutes (QSG‐60) or at 58°C for 60 minutes with proteinase‐K (QSG‐PK). Each HBV DNA sample was tested by heminested PCR amplification of the HBV gene sequences. A total of 36 coded serum samples were tested, including three HBsAg‐positive controls and 33 former chronic HBV carriers who had seroconverted (developed antibody to HBsAg [anti‐HBs]). Commercially available seroconversion panels (PHM 907, 911, and 922) were similarly tested for window‐period viremia.RESULTS:In the 33 former chronic HBV carriers who had seroconverted, IAC revealed HBV DNA in 17 samples, whereas it was revealed in only 11 samples by QSG‐PK (p = 0.031), 10 by QSG‐60 (p = 0.016), and 9 by QSG (p = 0.0078). However, HBV DNA was not amplified from the 17 samples at 1‐in‐10 dilutions; thus, they were considered to have low‐level viremia. IAC revealed HBV DNA as early as or earlier than the other methods in PHM 907, 911, and 922 panels.CONCLUSION:IAC is apparently an optimal method of sample preparation for amplification of HBV DNA in patients in the pre‐HBsAg window period, and for detecting low‐level viremia persistent in several individuals who were former chronic HBV carriers who had seroconverted (developed anti‐HBs).