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目的了解学龄前儿童(2~6岁)隐匿性乙型肝炎病毒感染(OBI)后HBV S基因氨基酸的变异情况,为防控OBI的发生发展提供参考。方法采用整群抽样方法选择乌鲁木齐4所幼儿园,对在园的学龄前儿童采用ELISA进行HBs Ag,anti-HBs,HBe Ag,anti-HBe和anti-HBc 5项检测,对259例HBs Ag阴性的血清用Promega公司生产的试剂盒进行HBV-DNA产物提取,运用PCR方法扩增HBV-DNA S基因片段,对扩增阳性的序列进行测定;采用分子遗传学软件MEGA5.0对测得的序列进行具体氨基酸分析。结果 259份学龄前儿童血样共发现21份HBV DNA阳性标本,均测序成功。其中1例由于存在氨基酸的插入,没有检出具体的基因型。其余20例样本均表现为C基因型,血清型均为adrq+亚型。S基因序列翻译成氨基酸后分别发生I110L,S123T,T124S,T126I,T131N,F134Y和P142H突变。结论 HBs Ag阴性学生血液中存在一定数量的HBV隐匿性感染,S基因存在氨基酸突变,需重视隐匿性的HBV感染以及其发生和发展。
Objective To understand the variation of amino acid in HBV S gene after occult hepatitis B virus infection (OBI) in preschool children (2 ~ 6 years old), and to provide a reference for preventing and controlling the occurrence and development of OBI. Methods A total of 4 kindergartens in Urumqi were selected by cluster sampling method. Five preschool children in the park were tested for HBsAg, anti-HBs, HBeAg, anti-HBe and anti-HBc using ELISA. 259 HBsAg-negative The serum was extracted with the kit manufactured by Promega Company, and the HBV-DNA S gene fragment was amplified by PCR, and the sequence of the positive amplification was determined. The measured sequence was performed by molecular genetic software MEGA5.0 Specific amino acid analysis. Results Twenty-one positive samples of HBV DNA were found in 259 preschool children’s blood samples, all of which were successfully sequenced. One of the cases did not detect a specific genotype due to the presence of amino acid insertion. The remaining 20 samples showed C genotype, serotypes are adrq + subtype. The S110L, S123T, T124S, T126I, T131N, F134Y and P142H mutations occur after the S gene sequence is translated into amino acids. Conclusions There is a certain amount of occult HBV infection in the blood of HBsAg-negative students. There is a mutation of amino acid in S gene. It is necessary to pay attention to occult HBV infection and its occurrence and development.