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目的探讨多细胞株微量培养法在病毒性脑炎、脑膜炎病原学检测中的应用价值。方法将脑脊液(CSF)标本过滤除菌后分别接种于含有青霉素和链霉素微量细胞培养板单层细胞上,每份标本平行接种8株细胞:MA104、MDCK、HEL、HEK293、Vero、Hep-2、Hela和BHK-21。静置培养,获得稳定的病毒株。以肠道病毒(EV)组合血清鉴定引起病毒性脑炎、脑膜炎的常见病毒类型。结果 126份病毒性脑炎、脑膜炎患儿CSF标本82份分离到病毒,阳性率63.0%,其中MA104阳性率为57.1%,MDCK阳性率为19.8%,HEL阳性率为11.9%,HEK阳性率为7.1%,Vero阳性率为6.3%,Hep-2阳性率为4.8%,Hela阳性率为4.0%,BHK-21阳性率为2.4%。病变细胞经血清学鉴定,78例鉴定出病毒株,鉴定率为92.8%,这些病毒中EV所占比例较高,为69.2%;ADV占11.5%,HSV-Ⅰ占5.1%,HSV-Ⅱ占6.4%;CMV占5.1%,INF占2.6%。结论 MA104细胞是分离CSF中病毒较敏感的细胞,可以作CSF中病原分离的首选细胞;联合多株细胞可提高细胞病变检出率。多株细胞微孔板培养法分离CSF病毒具有推广应用价值。
Objective To investigate the value of multi-cell culture in the etiological detection of viral encephalitis and meningitis. Methods CSF samples were inoculated into monolayer cells containing penicillin and streptomycin microtiter plate culture respectively. Eight samples were inoculated into each of them in parallel: MA104, MDCK, HEL, HEK293, Vero, Hep- 2, Hela and BHK-21. Static culture, to obtain a stable virus strain. The combination of enterovirus (EV) sera to identify viral encephalitis, meningitis common virus types. Results Among 82 viral encephalitis and meningitis children, 82 were isolated from CSF samples, the positive rate was 63.0%. The positive rate of MA104 was 57.1%, the positive rate of MDCK was 19.8%, the positive rate of HEL was 11.9% Was 7.1%. The positive rate of Vero was 6.3%, the positive rate of Hep-2 was 4.8%, the positive rate of Hela was 4.0% and the positive rate of BHK-21 was 2.4%. The pathological cells were identified by serology and 78 strains were identified. The identification rate was 92.8%. The proportion of EV in these viruses was high (69.2%). ADV was 11.5%, HSV-Ⅰ was 5.1%, and HSV-Ⅱ was 6.4%, CMV 5.1% and INF 2.6%. Conclusion MA104 cells are more sensitive to virus isolation in CSF and can be the first choice for pathogen isolation in CSF. Combined with multiple cells, the detection rate of cytopathic effect can be increased. Multi-strain cell microplate culture separation of CSF virus has the promotion and application value.