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首页> 外文期刊>Journal of Medical Entomology >Blinded laboratory comparison of the in situ enzyme immunoassay, the VecTest wicking assay, and a reverse transcription-polymerase chain reaction assay to detect mosquitoes infected with West Nile and St. Louis encephalitis viruses
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Blinded laboratory comparison of the in situ enzyme immunoassay, the VecTest wicking assay, and a reverse transcription-polymerase chain reaction assay to detect mosquitoes infected with West Nile and St. Louis encephalitis viruses

机译：原位酶免疫测定，VecTest芯吸测定和逆转录聚合酶链反应测定的盲实验室比较，以检测感染了西尼罗河和圣路易斯脑炎病毒的蚊子

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摘要

A blinded laboratory evaluation compared the accuracy, sensitivity, and specificity of an in situ enzyme immunoassay (EIA), VecTest wicking assay, and reverse transcription-polymerase chain reaction (RT-PCR) to detect and distinguish West Nile (WN) and St. Louis encephalitis (SLE) viruses in pools of 50 mosquitoes. Adult female Culex tarsalis Coquillett were inoculated with either WN or SLE viruses, held for 0-11 d at 28 degrees C, killed by freezing, and then were added to 49 or 48 uninfected mosquitoes to make up 14 pools positive for WN virus, 14 positive for SLE virus, 14 positive for both WN and SLE viruses, and 14 negative for virus. Pools were number coded and tested blindly. Virus was not detected in known negative pools. VecTest and RT-PCR assays were comparably sensitive and accurate, detecting virus in pools containing females held for 3 d postinoculation; only RT-PCR detected SLE virus in pools on days 0-1. The VecTest and RT-PCR produced a single false-positive result for WN and SLE, respectively. RT-PCR detected RNA in samples positive by the VecTest, indicating that the detergent in the wicking buffer did not prevent RT-PCR from confirming VecTest results. Detector antibodies used in the in situ EIA cross-reacted between SLE and WN viruses, reducing accuracy. Both the VecTest and RT-PCR provided rapid and specific results, but they detected only those viruses known to be present. Plaque assay on Vero cells was comparably sensitive and had the added benefit of detecting newly emerging viruses, but this method required virus culture followed by identification, thereby delaying reporting.
机译：盲目的实验室评估比较了原位酶免疫测定（EIA），VecTest芯吸测定和逆转录聚合酶链反应（RT-PCR）的准确性，敏感性和特异性，以检测和区分西尼罗河（WN）和圣尼罗河。每50只蚊子中有路易脑炎（SLE）病毒。用WN或SLE病毒对成年雌性库蚊（Culex tarsalis Coquillett）进行接种，在28摄氏度下保持0-11天，冷冻杀死，然后添加到49或48个未感染的蚊子中，组成14个WN病毒阳性的库，14 SLE病毒呈阳性，WN和SLE病毒均为14呈阳性，病毒为14阴性。对池进行数字编码并盲目测试。在已知的阴性池中未检测到病毒。 VecTest和RT-PCR分析具有相当的敏感性和准确性，可在含有雌性疫苗接种后3 d的池中检测病毒。在第0-1天，只有RT-PCR在池中检测到SLE病毒。 VecTest和RT-PCR分别对WN和SLE产生单个假阳性结果。 RT-PCR检测到VecTest呈阳性的样品中的RNA，表明芯吸缓冲液中的去污剂不会阻止RT-PCR确认VecTest结果。原位EIA中使用的检测器抗体在SLE和WN病毒之间发生交叉反应，从而降低了准确性。 VecTest和RT-PCR均提供了快速而特定的结果，但它们仅检测到已知存在的那些病毒。在Vero细胞上进行噬菌斑测定具有相当的敏感性，并具有检测新出现的病毒的额外好处，但是这种方法需要进行病毒培养，然后进行鉴定，从而延迟了报告。

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