基于PMA-qPCR方法检测兽用消毒剂对非洲猪瘟病毒的灭活效率

doi:10.3969/j.issn.1004-1524.2023.02.07

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (2): 301-307.DOI: 10.3969/j.issn.1004-1524.2023.02.07

基于PMA-qPCR方法检测兽用消毒剂对非洲猪瘟病毒的灭活效率

杨悦¹(), 邵靓², 张鹏飞¹, 陈弟诗², 陈婉婷¹, 王印¹^,³^,^*(), 罗燕¹^,³, 杨泽晓¹^,³, 姚学萍¹^,³, 任梅渗¹^,³

1.四川农业大学动物医学院,四川成都 611130
2.四川省动物疫病预防控制中心,四川成都 611130
3.动物疫病与人类健康四川省重点实验室,四川成都 611130

收稿日期:2022-04-01 出版日期:2023-02-25 发布日期:2023-03-14
通讯作者: 王印
作者简介:*王印,E-mail:yaanwangyin@tom.com
杨悦(1997—),女,四川成都人,硕士研究生,主要从事动物传染病病原分子生物学研究。E-mail:348265376@qq.com
基金资助:
四川省科技计划(2021YJ0270);四川省“十四五”川猪重大科技专项(2021ZDZX0010)

Inactivation efficiency of various disinfectants against African swine fever virus evaluated by PMA-qPCR

YANG Yue¹(), SHAO Jing², ZHANG Pengfei¹, CHEN Dishi², CHEN Wanting¹, WANG Yin¹^,³^,^*(), LUO Yan¹^,³, YANG Zexiao¹^,³, YAO Xueping¹^,³, REN Meishen¹^,³

1.College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
2. Sichuan Animal Disease Prevention and Control Center, Chengdu 611130, China
3. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, China

Received:2022-04-01 Online:2023-02-25 Published:2023-03-14
Contact: WANG Yin

摘要/Abstract

摘要：

为评估规模化猪场生物安全防控水平,拟建立非洲猪瘟病毒(African swine fever virus, ASFV)感染性检测方法,并利用该方法检测兽用消毒剂对病毒的灭活效果。使用不同种类的消毒剂处理ASFV,通过PMA-qPCR评估消毒剂对ASFV的灭活效果和筛选消毒剂的最佳处理方式。PMA-qPCR结果显示,ASFV对不同类型的消毒剂表现出不同的敏感性,对过硫酸氢钾复合物溶液敏感性最高,该消毒剂按0.5%稀释,20 min内即可有效灭活ASFV。研究结果对生物安全的消毒环节具有一定的参考意义。

关键词: 非洲猪瘟病毒, 消毒剂, 叠氮溴化丙锭, 生物安全

Abstract:

In order to evaluate the biosecurity prevention and control level of large-scale pig farms, this study intends to establish a method to detect the infectivity of African swine fever virus(ASFV), and use this method to detect the inactivation effect of various disinfectants on ASFV. We established a method called PMA-qPCR, which was used to evaluate the inactivation effect after treating ASFV with different kinds of disinfectants, and used it to explore the best treatment method of disinfectants. The results of PMA-qPCR showed that ASFV had different sensitivities to different types of disinfectants, with the highest sensitivity to potassium hydrogen persulfate complex solution, which could effectively inactivate ASFV within 20 min after diluting at 0.5%. The results had certain reference for the disinfection of biosecurity.

Key words: African swine fever virus, disinfectant, propidium monoazide, biosecurity

中图分类号:

S851.36

杨悦, 邵靓, 张鹏飞, 陈弟诗, 陈婉婷, 王印, 罗燕, 杨泽晓, 姚学萍, 任梅渗. 基于PMA-qPCR方法检测兽用消毒剂对非洲猪瘟病毒的灭活效率[J]. 浙江农业学报, 2023, 35(2): 301-307.

YANG Yue, SHAO Jing, ZHANG Pengfei, CHEN Dishi, CHEN Wanting, WANG Yin, LUO Yan, YANG Zexiao, YAO Xueping, REN Meishen. Inactivation efficiency of various disinfectants against African swine fever virus evaluated by PMA-qPCR[J]. Acta Agriculturae Zhejiangensis, 2023, 35(2): 301-307.

图/表 8

参考文献 18

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名称Name	序列Sequence (5'→3')
ASFV-F	CTGCTCATGGTATCAATCTTATCGA
ASFV-R	GATACCACAAGATCAGGCCGT
ASFV-P	FAM-CCACGGGAGGAATACCAACCCAGTG-TAMRA

名称Name	序列Sequence (5'→3')
ASFV-F	CTGCTCATGGTATCAATCTTATCGA
ASFV-R	GATACCACAAGATCAGGCCGT
ASFV-P	FAM-CCACGGGAGGAATACCAACCCAGTG-TAMRA

组别 Group	处理方式 Treatment
1	高压蒸汽灭菌(103.4 kPa,121℃),添加PMA Autoclaved (103.4 kPa,121 ℃)+PMA
2	常温处理,添加PMA Room temperature treatment+PMA
3	高压蒸汽灭菌(103.4 kPa,121℃),不添加PMA Autoclaved (103.4 kPa,121 ℃)
4	常温处理,不添加PMA Room temperature treatment

组别 Group	处理方式 Treatment
1	高压蒸汽灭菌(103.4 kPa,121℃),添加PMA Autoclaved (103.4 kPa,121 ℃)+PMA
2	常温处理,添加PMA Room temperature treatment+PMA
3	高压蒸汽灭菌(103.4 kPa,121℃),不添加PMA Autoclaved (103.4 kPa,121 ℃)
4	常温处理,不添加PMA Room temperature treatment

组别 Group	处理方式 Treatment	Ct值 Ct value
1	高压蒸汽灭菌(103.4 kPa,121℃),添加PMA Autoclaved(103.4 kPa,121 ℃)+PMA	>40
2	常温处理,添加PMA Room temperature treatment+PMA	31.03
3	高压蒸汽灭菌(103.4 kPa,121℃),不添加PMA Autoclaved (103.4 kPa, 121 ℃)	32.27
4	常温处理,不添加PMA Room temperature treatment	30.69

基于PMA-qPCR方法检测兽用消毒剂对非洲猪瘟病毒的灭活效率

Inactivation efficiency of various disinfectants against African swine fever virus evaluated by PMA-qPCR

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组别 Groups	消毒剂稀释度与种类 Concentrations and types of disinfectants	灭活率 Inactivation rate/%
1	0.5%过硫酸氢钾复合物 0.5% Potassium hydrogen persulfate complex	99.87
2	1%二氯异氰脲酸钠 1% Sodium dichloroisocyanurate	98.14
3	2.5%二氧化氯 2.5% Chlorine dioxide	97.46
4	0.2%复合碘 0.2% Compound iodine	96.45
5	0.5%辛硫磷0.5% Phoxim	81.75
6	0.2%复合戊二醛季铵盐溶液 0.2% Compound glutaraldehyde quaternary ammonium salt solution	73.51
7	1%有机酸 1% Organic acids	67.83
8	0.5%溴化二甲基二癸基羟胺 0.5% Dimethyl didecyl hydroxylamine bromide	65.44
9	0.25%戊二醛-癸甲溴铵 0.25% Glutaraldehyde decamethonium bromide	62.84

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