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目的调查2009—2011年辽宁省8市发热伴血小板减少综合征新病原,探讨新布尼亚病毒在自然界中可能存在的动物宿主及传播媒介。方法采用ELISA方法检测2009—2011年辽宁省发热伴血小板减少患者血液中发热伴血小板减少综合征布尼亚病毒(SFTSV)IgM抗体,采用间接免疫荧光染色法检测鼠肺SFTSV抗原,利用逆转录-聚合酶链反应(RT-PCR)检测鼠肺、蜱悬液、细胞阳性分离物的SFTSV核酸,采用VERO-E6细胞对患者血液及脾悬液标本进行病毒分离鉴定,并进行核苷酸测序分析。结果大连、丹东、抚顺、铁岭均检出SFTSV的IgM抗体阳性;103份中检出20份鼠肺SFTSV抗原阳性,但SFTSV核酸检测均阴性;20组中1组蜱悬液SFTSV核酸检测阳性并分离出SFTSV病毒;148份中有26份急性期血液样本经VERO-E6细胞分离出现细胞病变,经鉴定为SFTSV病毒,并对M片段进行核苷酸序列分析。结论新布尼亚病毒是辽宁省4市发热伴血小板减少综合征的新病原,未检出可能的自然界中的动物宿主鼠类携带SFTSV核酸,蜱可能是其传播媒介之一。
Objective To investigate the new pathogens of fever with thrombocytopenic syndrome in 8 cities of Liaoning Province from 2009 to 2011 and to explore the possible animal hosts and vectors of New Bunyan virus in nature. Methods The serum anti-SFTSV IgM antibodies against fever and thrombocytopenia syndrome were detected by enzyme-linked immunosorbent assay (ELISA) in Liaoning Province from 1999 to 2011. The SFTSV antigens were detected by indirect immunofluorescence staining. The SFTSV nucleic acids of lung, tick suspension and cell positive isolates were detected by polymerase chain reaction (RT-PCR). VERO-E6 cells were used for virus isolation and identification of the blood and spleen suspension samples. Nucleotide sequencing . Results SFTSV IgM antibody positive was detected in Dalian, Dandong, Fushun and Tieling. Totally 20 SFTSV antigens were detected in 103 mice, but negative in SFTSV nucleic acid test. SFTSV nucleic acid was detected in 1 group of tick suspension in 20 groups SFTSV virus was isolated. Among the 148 samples, 26 acute phase blood samples were isolated from VERO-E6 cells and identified as SFTSV virus. The M fragment was sequenced. Conclusion New Bunyan virus is a new pathogen of fever with thrombocytopenia syndrome in 4 cities of Liaoning Province. It is not possible to detect SFTSV nucleic acid in animal hosts that may be found in nature. Tick may be one of its vectors.