脊髓灰质炎病毒细胞系敏感性实验方法的建立

Establishment of Cell Sensitivity Test for Poliovirus

目的评价中国疾病预防控制中心(CDC)病毒病预防控制所国家脊髓灰质炎(脊灰)实验室所用细胞系对脊灰病毒的敏感性,制备国家脊灰实验室质量控制株(China Sabin Master Reference,CSMR)和用于脊灰实验室网络的中国Sabin参考株(China Sabin Test Reference Strain;CSTRS),提供给省级CDC脊灰实验室用于其对所使用的RD-A、L20B细胞系的敏感性评价。方法96孔微量培养板滴定法。结果国家脊灰实验室CSMR 3次独立的细胞敏感性实验结果的滴度波动在±0.5log10,同时用英国国家生物标准品与控制样品研究所(NIBSC)提供的已知滴度Sabin脊灰病毒参考株(NIBSC原始株)作平行对照,NIBSC原始株3次滴度结果与其本身提供的参考值相比较,其滴度波动也均在±0.5log10。结论通过NIBSC原始株3次滴度结果作平行对照,证实中国CSMS结果符合实验要求,国家脊灰实验室的内部CSMR结果有效;从而制定了国家脊灰实验室质量控制参考株的滴度参考值,表明国家脊灰实验室所用细胞系的敏感性未下降,实验程序无误。CSTRS可作为标准株分发到省级CDC脊灰实验室用于进行细胞系敏感性实验。

Objective To evaluate the sensitivity of cell lines for poliovirus isolation in China National Polio Laboratory, make the China Sabin Master Reference （CSMR）, and China Sabin Test Reference Strain （CSTR） for the whole laboratory network in China, which can be supplied to 31 provincial polio laboratories to test the sensitivity of the RD-A and L20B cell lines. Method 96-microwells plates culture and titration. Result In China, national polio laboratory, we have finished the sensitivity test of CSMR, after three times individual tests,we found that the results vary in ＋ 0.5log10, at the same time, we used the Sabin reference strain from British National Institute of Biological Standard and Control （NIBSC） as parallel tests,the sensitivity test of the NIBSC strain were also vary ＋ 0.5log10 within 3 individuals tests comparing with its standard. Conclusion The cell sensitivity test have been established by the China National Laboratory for Poliomyelitis （CNLP） . Through three times parallel tests, we can confirm that China CSMR accord with the demand, the internal CSMR of CNLP is valid, we have made the titration reference of CSMR to be the national laboratory quality control, all the facts have shown that the sensitivity of the RD-A and L20B lines used in CNLP is not dropping, the testing procedure is correct. The CSTR, as a standard strain, has been distributed to the provincial polio laboratories network as the reference strain of the cell lines sensitivity tests.