猴B病毒血清学和PCR检测结果比较

Comparison of Monkey B Virus Determination with Serological Assays and PCR Test

目的:比较猴B病毒血清抗体和病毒PCR检测结果,阐明动物感染后病毒在机体内的存在状况。方法:采集成年猴血清和三叉神经组织,首先通过ELISA方法检测血清中B病毒抗体,然后采用B病毒gL和gD基因引物通过PCR方法扩增血清DNA和三叉神经组织DNA,比较2种方法的检测结果,并对扩增产物进行序列分析。结果:22份猴血清中,B病毒抗体呈阳性的有13份(59.1%);PCR结果显示,抗体阴性动物及所有血清DNA模板中均无阳性扩增,但在13份抗体阳性动物的三叉神经组织DNA样品中,PCR阳性4份(30.8%);gL和gD基因扩增条件及产物分析表明,gL基因的GC含量为64.1%,gD为74.2%,且gL的扩增条件和效果明显优于gD。结论:B病毒感染猴后,将在部分动物神经节中建立潜伏,而gL基因更适合作为分子鉴定的靶标。

Objective： To elucidate the state of monkey B virus（BV） in infected monkey, antibodies and viral DNA in 22 monkeys were detected at the same time. Methods： Sera and trigeminal ganglia（TG） tissue were col- lected from 22 adult monkeys. At first, antibodies to BV in sera were detected by dot immunobinding assay （DIA）. Then, DNA was extracted and purified from sera and TG tissue and used as templates for PCR with prim- ers on gL or gD gene of BV. Products of PCR were sequenced and blast in GenBank. Results： In 22 monkey se- ra, 13（59.1%） were confirmed for the presence of antibodies to BV. Results of PCR showed that the virus DNA was present in TG of 4 of the 13 seropositve monkeys（30.8%） tested, but no virus being detected in all sera sam- ples. Sequencing products of PCR showed G＋C content in gD gene was very high（74.2%） and middle in gL gene （64.1%）. The target of gL gene for PCR was superior to gD gene. Conclusion： Our results indicated that B vi- rus would establish latent in tfigeminal ganglia only in part of infected moneys and gL gene locus of BV is a suit- able target for specific and rapid identification of viral infection by PCR technology.