三种不同灭活剂对猪细小病毒和猪圆环病毒的灭活效果研究

Inactivation effects of 3 different inactivators on porcine parvovirus and porcine circovirus

使用不同浓度的甲醛、β-丙内酯(BPL)、二乙烯亚胺(BEI)对猪细小病毒(S-1株)、猪圆环病毒2型(JSM株)进行灭活处理,采用灭活检验试验[血凝(HA)试验和IFA试验]和无菌检验试验检测不同时间段采集的灭活样品,研究不同灭活剂对猪细小病毒和猪圆环病毒的灭活效果,筛选最佳灭活条件。结果表明:盲传3代后,在37℃条件下,0.2%的甲醛灭活48 h、36 h可彻底灭活S-1株和JSM株;在32℃条件下,BEI终浓度为0.1%时,对S-1株和JSM株的最佳灭活时间分别是84 h和60 h;在4℃条件下,BPL对S-1株和JSM株的灭活终浓度分别为0.03%、0.05%,灭活时间分别为72 h、48 h。HA试验结果显示:使用甲醛灭活后,猪细小病毒血凝效价由2^(11)下降至2~6—2~5;使用BPL与BEI灭活对PPV血凝活性无明显影响。灭活终止程序中,BEI需要添加总体积10%的硫代硫酸钠,BPL在37℃、2 h即可完全水解。本研究最佳灭活条件为:在4℃条件下,终浓度为0.05%的BPL灭活48 h。

Different concentrations of formaldehyde,β-propiolactone( BPL) and diethyleneimine( BEI)were used to inactivate PPV(S-1 strain) and PCV2(JSM strain).The inactivation test[HA test and IFA test]and sterility test were used to detect the inactivation samples collected at different time to study the inactivation effects of different inactivators on porcine parvovirus and porcine circovirus,and to screen the best inactivation conditions.The results showed that S-1 strain and JSM strain could be completely inactivated by 0.2%formaldehyde for 48 h and 36 h at 37 ℃ after blind passage 3 generations;At 32 ℃,when the final concentration of BEI was 0.1%,the best inactivation time for S-1 strain and JSM strain was 84 h and 60 h respectively;At 4 ℃,the final inactivation concentration of BPL for S-1 strain and JSM strain was 0.03% and0.05%,respectively,and the inactivation time was 72 h and 48 h,respectively.HA test results showed that the hemagglutination titer of porcine parvovirus decreased from 2^(11) to 2~6—2~5 after inactivation with formaldehyde;Inactivation with BPL and BEI had no significant effect on the hemagglutination activity of PPV.In the inactivation termination procedure,BEI needed to add 10% of the total volume of sodium thiosulfate,and BPL could be completely hydrolyzed at 37 ℃ for 2 h.The optimal inactivation conditions in this study were inactivation with 0.05% BPL at 4 ℃ for 48 h.