CAJ | 학술논문

目的探讨肠道病毒VP4基因通用引物PCR诊断非肠道病毒71型( EV71)非柯萨奇病毒A组16型( CoxA16)手足口病( HFMD)病毒方案的准确性。方法通过肠道病毒VP4基因通用引物PCR鉴定阜阳地区非EV71非CoxA16 HFMD病毒时,显示两例病毒分离株为Sabin-1(命名为Fy-01和Fy-02株)。分别采用引物EVP4/Q8和VP1特异性引物对Fy-01和Fy-02分离株进行鉴定。结果采用VP4基因PCR扩增产物测序证实,两例病毒分离株为Sabin-1,而采用EVP4/Q8引物PCR扩增Fy-01分离株却存在异常的扩增条带,PCR扩增产物测序结果为柯萨奇病毒A组10型( Cox-A10)；将VP4基因PCR产物制备成克隆载体挑选多个单克隆测序结果显示,Fy-01分离株既存在Sabin-1 VP4基因,又存在CoxA10 VP4基因,证实为两种毒株混合感染。结论临床诊断为非EV71非CoxA16 HFMD的病例,如病原学诊断仅采用VP4基因PCR产物测序,特别是针对多种肠道病毒混合感染引起的HFMD,其结果并不完全可靠。
목적탐토장도병독VP4기인통용인물PCR진단비장도병독71형( EV71)비가살기병독A조16형( CoxA16)수족구병( HFMD)병독방안적준학성。방법통과장도병독VP4기인통용인물PCR감정부양지구비EV71비CoxA16 HFMD병독시,현시량례병독분리주위Sabin-1(명명위Fy-01화Fy-02주)。분별채용인물EVP4/Q8화VP1특이성인물대Fy-01화Fy-02분리주진행감정。결과채용VP4기인PCR확증산물측서증실,량례병독분리주위Sabin-1,이채용EVP4/Q8인물PCR확증Fy-01분리주각존재이상적확증조대,PCR확증산물측서결과위가살기병독A조10형( Cox-A10)；장VP4기인PCR산물제비성극륭재체도선다개단극륭측서결과현시,Fy-01분리주기존재Sabin-1 VP4기인,우존재CoxA10 VP4기인,증실위량충독주혼합감염。결론림상진단위비EV71비CoxA16 HFMD적병례,여병원학진단부채용VP4기인PCR산물측서,특별시침대다충장도병독혼합감염인기적HFMD,기결과병불완전가고。
Objective To investigate the accuracy of PCR for detection of non-EV71 and non-CoxA16 by using u-niversal primers with VP4 gene of HEV. Methods When identified by VP4 gene of EV71 and CoxA16 unrelated HFMD, two isolates of Sabin-1 were found(such as Fy-01and Fy-02). They were identified by co-primers such as EVP4/Q8 and specific primers of VP1 gene. Results Fy-01 isolates were appeared with an unusual frayment and identified as CoxA10 after being sequenced and the selected independent clones were sequenced and blasted in GenBank. It proved that both Sabin-1 and CoxA10 existed. Conclusion Clinical diagnosis of EV71 and CoxA16 unrelated HFMD,when identifying the type of HFMD pathogen only by using the amplification product of VP4 gene after being sequenced directly is not enough, especially HFMD with mixed infections caused by enterovirus.