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目的了解广东省肇庆市2014-2015年登革病毒(DENV)的基因型别和可能的输入来源。方法收集登革热临床诊断病例急性期血清,对实时荧光反转录-聚合酶链反应(RT-PCR)检测阳性标本进行病毒培养,分离株进行E基因扩增与序列测定,用Mega 5.1软件进行分子分型和系统进化树构建。结果 409份样品中147份实时荧光RT-PCR检测阳性(35.94%),其中2014年阳性144份(39.99%),以DENV 1(139/361)为主;2015年阳性3份(6.25%),以DENV 3(2/48)为主。共获得22株DENV分离株,其中20株为2014年DENV 1型的genotypeⅠ和Ⅲ,与2013-2014年广州、佛山市分离株核苷酸同源性分别为99.40%~100.00%和99.20%~100.00%,与新加坡、泰国、马来西亚和印度尼西亚分离株亲缘关系次之;2株为2015年DENV 3的genotypeⅢ,与近年新加坡和印度尼西亚分离株的核苷酸同源性为98.20%~99.70%。结论广东省肇庆市2014年暴发流行的DENV 1为genotypeⅠ和Ⅲ,可能由广州市和佛山市输入,2015年散发的DENV 3为genotypeⅢ,由佛山市输入,疫情毒株可能源于新加坡、印度尼西亚等东南亚流行株。
Objective To understand genotypes and possible sources of dengue virus (DENV) in Zhaoqing City, Guangdong Province during 2014-2015. Methods Serum was collected from the acute phase of clinical diagnosis of dengue fever. The positive samples were detected by real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR), and the E genes were amplified and sequenced. Mega 5.1 software Classification and phylogenetic tree construction. Results 147 of the 409 samples were positive by RT-PCR (35.94%), of which 144 were positive (39.99%) in 2014, and were mainly DENV 1 (139/361). In 2015, 3 were positive (6.25% , Mainly DENV 3 (2/48). A total of 22 DENV isolates were obtained, of which 20 were genotypes I and III of DENV type 1 in 2014, and 99.40% -100.00% and 99.20% respectively of isolates from Guangzhou and Foshan in 2013-2014, 100.00%, followed by Singapore, Thailand, Malaysia and Indonesia isolates. Two strains were genotype Ⅲ of DENV 3 in 2015, which shared 98.20% -99.70% nucleotide similarity with Singapore and Indonesia in recent years. Conclusions The outbreaks of DENV 1 in Zhaoqing City of Guangdong Province in 2014 were genotype Ⅰ and Ⅲ, possibly imported from Guangzhou and Foshan. In 2015, DENV 3 was genotype Ⅲ, imported from Foshan. The outbreak strains may originate from Singapore, Indonesia, etc. Southeast Asia epidemic strains.