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目的对云南地区狂犬病疫情开展分子流行病学调查与分析。方法应用直接免疫荧光试验(DFA)检测动物和患者中的狂犬病毒抗原,阳性样本用反转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)扩增狂犬病毒N基因片段,并进行序列测定及分析。结果对60份患病动物样本进行DFA检测,结果抗原检测阳性26份(阳性率43.33%)。对182份健康犬脑进行DFA检测,结果抗原检测阳性2份(阳性率1.10%)。对4份患者唾液和73份鼠脑进行DFA检测,抗原检测均为阴性。对28份抗原检测阳性样本进行RT-PCR检测,25份核酸检测阳性(阳性率89.29%)。基因进化树分析显示本次检测到的DX01和DZ01病毒与国内狂犬病毒株在同一分支之内,DN01与东南亚毒株在同一分支之内。结论引发云南省狂犬病流行的狂犬病毒较复杂,其来源可能与东南亚和省外毒株的输入有关。
Objective To carry out molecular epidemiological investigation and analysis of rabies epidemic in Yunnan Province. Methods The rabies virus antigens were detected by direct immunofluorescence assay (DFA) in rabbits and patients. The positive samples were amplified by reverse transcription-polymerase chain reaction (RT-PCR) And sequence determination and analysis. Results DFA was detected in 60 diseased animal samples. As a result, 26 positive samples were detected (positive rate 43.33%). 182 healthy dogs brain DFA test, the results of positive antigen test 2 (positive rate 1.10%). DFA test was performed on 4 patients’ saliva and 73 mouse brain, and the antigen test was negative. Twenty-eight positive samples were tested by RT-PCR and 25 were positive (89.29% positive). Phylogenetic tree analysis showed that the DX01 and DZ01 viruses detected in this study were in the same branch with the domestic rabies virus strains, and DN01 was in the same branch with the strains in Southeast Asia. Conclusions The rabies virus that caused the rabies epidemic in Yunnan Province is more complicated and its origin may be related to the input of strains in Southeast Asia and other provinces.