青岛市首例新型布尼亚病毒L基因全长分析

Analysis of specific L fragments about the first novel Bunyavirus in Qingdao

目的应用real-time PCR对青岛市首例发热伴血小板减少综合征患者血清的新型布尼亚病毒进行扩增,对病毒L基因测序及序列比对技术进行分析鉴定。方法选取青岛市首例发热伴血小板减少综合征患者急性期血清1份,利用荧光RT-PCR进行测定为阳性,利用套式RT-PCR进行L特异性基因片段全长扩增并进行测序拼接,结果与Gen Bank公布的序列进行比对确认。用DNAStar软件构建进化树进行分子流行病学研究。结果青岛市首例发热伴血小板减少综合征L基因全长与江苏2014年毒株KR23076的同源性为99. 7%,与山东2010年的毒株JQ684871同源性为99. 8%。结论青岛市首例发热伴血小板减少综合征L基因序列全长鉴定成功,为青岛市该疾病研究和监测提供数据支持。

Objective Real-time PCR was used to amplify the novel Bunyavirus in the serum of patients with fever and thrombocytopenia syndrome in Qingdao. The virus L gene was sequenced and analyzed by sequence alignment technique.Methods The acute stage serum of the first patient with fever and thrombocytopenia syndrome in Qingdao was selected and detected as positive by fluorescent RT-PCR. The full-length amplification of L-specific gene fragments was performed by nested RT-PCR and sequenced. The result was compared with the sequence published by Gen Bank. The phylogenetic tree was constructed using DNAStar for molecular epidemiological studies. Results The full length of the L gene of the first fever with thrombocytopenia syndrome in Qingdao was 99. 7% homological with KR23076 in Jiangsu in 2014,and 99. 8% homological with JQ684871 in Shandong in 2010. Conclusion In Qingdao,the L gene sequence was successfully identified in the first fever with platelet reduction syndrome,which provided data support for the research and monitoring of the disease in Qingdao.