实时荧光定量PCR技术在手足口病病原体检测中的临床应用

Clinical application of real-time fluorescence quantification PCR technology in HFMD pathogen detection

目的：探讨实时荧光定量PCR（FQ-PCR）技术在手足口病（HFMD）病原体检测中的应用，了解遵义市HFMD流行特征。方法收集2015年5~7月该院儿科初步诊断为HFMD的85例患儿的咽拭子、疱疹液标本，采用FQ-PCR检测标本中肠道通用型病毒（EV-U）、肠道病毒71型（EV71）及柯萨奇病毒A组16型（CoxA16），同时与酶联免疫吸附试验（ELISA）检测结果进行比较分析。结果85例HFMD患儿采用FQ-PCR法检出65例EV-U阳性，阳性率为76.5％，其中2岁以下年龄段患儿阳性40例，阳性率为80.0%（40/50），2～5岁患儿阳性25例，阳性率为71.4%（25/35）。ELISA法检测EV-U阳性52例，其中2岁以下年龄段患儿阳性31例，阳性率为62.0%（31/50），2～5岁年龄段患儿阳性21例，阳性率为60.0％（21/35）。2种方法检测阳性率比较，差异有统计学意义（P〈0.05）。结论 FQ-PCR技术以其灵敏度高、特异性强、速率快等特点更适用于临床检测HFMD病原体。2015年遵义市HFMD病原体以EV71较多，其次是CoxA16。

Objective To investigate the application of real-time fluorescence quantitative polymerase chain reaction （FQ-PCR） technology in hand-foot and mouth disease（HFMD） pathogens detection,and to understand the HFMD epidemic char-acteristics in Zunyi City. Methods The throat swabs and herpes liquid samples were collected from 85 children patients with preliminarily diagnosed HFMD in the pediatric department of our hospital. EV-U,EV71 and coxsackievirus A 16（CoxA16） were detected by using the FQ-PCR technology,meanwhile the obtained results were compared with those detected by ELISA. Results Among 85 cases of HFMD,65 cases of EV-U positive were detected by FQ-PCR,the positive rate was 76.5%,in which 40 cases were in the age group of less than 2 years old, the positive rate was 80.0%（40/50）,25 cases were in the age group of 2-5 years, the positive rate was 71.4%（25/35）. Fifty-two cases of EV-U positive were detected by ELISA,in which 31 cases were the age group of under 2 years old,the positive rate was 62.0%（31/50）,21 cases were in the age group of 2-5 years,the positive rate was 60.0%（21/35）,the difference between these two kinds of method was statistically significant （P〈0.05）. Conclusion The FQ-PCR technology of HFMD pathogen detection is more suitable for the clinical application due to the characteristics of high sensitivity , strong specificity and high speed. The HFMD pathogens in Zunyi City during 2015 are mainly EV71 ,followed by CoxA 16.