水样中脊髓灰质炎病毒检测方法的实验研究

The experimental study on culturing polioviruses in environmental water with the methods of living or solidified L20B cell absorption

目的:探索灵敏、简便、实用的检测水样中微量脊髓灰质炎病毒(脊灰病毒)的方法.方法:将L20B细胞加入含微量脊灰病毒的水样中,充分作用后收集细胞,作病毒分离培养,并以NaCl-AlCl3沉淀法作对照.结果:应用活的L20B细胞时,1 000 ml水样中含病毒100TCID50、10TCID50时能全部检出;当含1～2TCID50时14份样本可检出13份.NaCl-AlCl3沉淀法在100TCID50时4份样本中仅检出1 份,其他各浓度的4份样未能检出;应用固相化的L20B细胞处理污水时,2～3×106个细胞,甲醛固定4 h和48 h以上的细胞吸附的病毒量,均值都是18.1TCID50,用细胞吸附浓集法,在40份自然水样中,10份检出脊灰病毒,而NaCl-AlCl3沉淀法未能检出脊灰病毒.结论:应用活细胞吸附时敏感度达到1～2TCID50/1 000ml;应用甲醛固定的细胞吸附污水中病毒时敏感度可达到10 TCID50/1 000ml.

Objective： To set up a easy, practical and sensitive methods for detecting minimum polio virus in large volume water. Methods： Adding L20B cells into the large volume water containing minimum polio virus. Collecting the cells after they fully reacted with viruses, then culturing viruses with the method of NaCl-AlCl3 sedimentation as control. Results： If the amount of viruses was 100 TCID50, 10TCID50 per 1000 ml water, the viruses could be detected with living cell absorption, and when the amount was 1-2TCID50, the viruses in 13 of 14 samples can also be detected. While the viruses of 100TCID50 per 1000ml water in 1 of 4 samples can be detected with the method of NaCl-AlCl3 sedimentation, but the viruses of other concentrations in all samples couldn＇t be detected. The average amount of absorbed polioviruses by 2-3×10^6 cells solidified with formaldehyde was 18.1TCID50. Of 40 natural water samples, tested with the method of cell absorption , the poliovirus were examined in 10 samples. Conclusions： The lowest absorption amount of living L20B cells was 1-2TCID50 per1000 ml water, and the lowest absorption amount of formaldehyde solidified cells was 10 TCID50 per1000ml water.