阜阳市手足口病病例巢式RT-PCR诊断与结果分析

Direct detection and result analysis of clinical specimens from a hand,foot,and mouth disease outbreak in Fuyang City by nested reverse transcription-PCR

目的明确阜阳市手足口病暴发的病原体,并对RT-PCR结果进行分析。方法利用巢式RT-PCR方法,分别采用肠道通用引物、EV71特异引物和CoxA16特异引物对这起暴发中156例病人的176份病例标本进行检测。结果156例病例的176份标本中,肠道通用引物、EV71特异引物和CoxA16特异引物检测阳性率分别为73.30%、63.07%、0%。咽拭子、气管吸痰液、肛拭子肠道通用引物检测阳性率分别为75.71%、54.54%、78.57%,χ2=4.57,P>0.05;EV71特异引物检测阳性率分别为65.00%、45.45%、71.43%,χ2=3.57,P>0.05,两种引物检测的三类标本阳性率比较均无统计学意义。156例病例中,有22人采集了双份或多份标本共计50份,其中标本阳性率和病例阳性率比较,肠道通用引物检测分别为56.00%、81.82%,χ2=4.41,P<0.05;EV71特异引物检测分别为54.00%、81.82%,χ2=5.04,P<0.05,两种引物检测结果均有统计学意义。结论巢式RT-PCR技术是肠道病毒快速诊断的重要手段,阜阳市手足口病暴发的病原体为肠道病毒EV71型。多次多样本采样能够提高病例阳性诊断率。

Objective To identify the etiological agents involved in the HFMD outbreak in Fuyang, and analyze the RT-PCR results. Methods The RT-PCR was used with universal pan-enterovirus primers, EV71 type-specific primers and coxsackievirus A16 type-specific primers for the direct detection of pathogen in 176 clinical specimens derived from this outbreak, Results Among 176 clinical specimens obtained from 156 patients suffering from HFMD in Fuyang, the positive rates of clinical specimens am- plified with universal primers, enterovirus 71 type-specific primers and coxsackievirus A16 type-specific primers were 73.30%, 63.07%, 0% respectively. The positive rates of throat swabs, tracheal aspi- rates and rectal swabs amplified with universal primers were 75.71%, 54.54%, 78.57%, χ^2=4.57, P〉0.05; amplified with EV71 type-specific primers were 65.00%, 45.45%, 71.43%, χ^2= 3.57, P 〉0.05. Three kinds of specimens amplified with two kinds of primers had no statistical significance. A total of 50 clinical specimens were obtained from 22 patients and two or more samples were collected from each patients. Respectively, amplified with universal primers, the positive rates of clinical specimens and patients were 56.00 %, 81.82 %, χ^2 = 4.41, P 〈 0.05 ; amplified with EV71 type-specific primers, the positive rates of clinical specimens and patients were 54.00%, 81.82%, χ^2 = 5.04, P〈 0.05, and had statistical significance amplified with two kinds of primers. Conclusions In view of its high sensitivity and specificity, nestsd reverse transeription-PCR is beneficial for the rapid detection of EV71 in outbreak situations. The major etiological agents of this HFMD outbreak in Fuyang are EV71. The positive rates of direct detection will be improved by multiple sample collection.