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目的探讨支原体污染对手足口病(Hand,Foot and Mouth Disease,HFMD)相关肠道病毒(Enterovirus,EV)分离率的影响。方法随机选择13个省的HFMD相关EV培养物共26份,同时从EV复核率较低的某省送检的HFMD相关EV培养物中,随机挑选18份,采用套式聚合酶链反应(Nested Polymerase Chain Reaction,nPCR)对细胞培养物中的支原体核酸进行检测。对阳性产物进行序列测定,通过基本多序列比对工具(Basic Local Alignment Search Tool,BLAST)对序列进行同源性比对。结果随机挑选13个省的EV培养物中,4个省的标本支原体检测结果为阴性,1个省同时检出猪鼻支原体和精氨酸支原体,8个省检出精氨酸支原体。EV复核率较低省送检的18份培养物中均检出牛支原体。结论支原体污染在中国HFMD实验室使用的细胞系中普遍存在,某些支原体的污染,可能降低细胞的敏感性,引起病毒分离率的下降。避免细胞系的支原体污染对提高HFMD实验室网络运转质量具有重要意义。
Objective To investigate the effect of mycoplasma contamination on the isolation rate of enterovirus (Enterovirus) in Hand, Foot and Mouth Disease (HFMD). Methods A total of 26 HFMD-related EV cultures were randomly selected from 13 provinces and 18 randomly selected HFMD-associated EV cultures from a province with a lower EV EV rate were analyzed by nested PCR Polymerase Chain Reaction, nPCR) Mycoplasma nucleic acids in cell cultures were tested. The positive products were sequenced and the sequences were aligned for homology using the Basic Local Alignment Search Tool (BLAST). Results Of the 13 EVs selected from 13 provinces, the Mycoplasma hyopneumoniae and Mycoplasma arbtans were detected negative in one of the four provinces. Mycoplasma arginsei was detected in 8 provinces. Mycoplasma bovidum was detected in 18 cultures tested in lower EV review rates. Conclusions Mycoplasma contamination is ubiquitous in cell lines used in HFMD laboratories in China. Contamination of some mycoplasma may reduce the sensitivity of cells and lead to the decrease of virus isolation rate. Avoiding mycoplasma contamination of cell lines is of great importance to improve the quality of operation of the HFMD laboratory network.