常见消毒剂、清洁剂对H5亚型禽流感病毒荧光RT-PCR检测结果的影响

The Influence of common disinfectants and detergents on the Test Results of H5 subtype avian influenza virus by the Fluorescent Quantitative RT-PCR

近期出现的H5N8亚型高致病性禽流感对家禽业构成了重大威胁,加强监测是做好禽流感防控的前提和基础。随着行业生物安全水平的提升,消毒剂在各生产流通环节得到较广泛的应用,但这些物质对当前监测使用的荧光RT⁃PCR方法检测结果的影响如何鲜见报道。本文采用禽流感病毒H5亚型HI试验抗原与相关干扰物质混合作为模拟样品,研究了生石灰、烧碱、二氯异氰酸钠和戊二醛等消毒剂,洗衣粉、洗洁精等清洁剂,甘油、头孢噻呋钠等样品保护剂,以及粪便等环境污物对H5亚型禽流感病毒核酸荧光RT⁃PCR检测结果的影响。结果显示常用消毒剂对检测结果影响极显著,与对照组相比,浓度高于0.5%时的生石灰处理组和浓度高于0.25%时的烧碱处理组出现不能检出情况,常规使用浓度的二氯异氰酸钠可导致荧光RT⁃PCR扩增产物的CT值增加10.29~10.61,戊二醛可致其CT值增加4.77~12.68。对含有碱性消毒剂的样品采取中和措施,可使检测结果有所改善。其他清洁剂、样品保护剂、污物对CT值均有一定的影响,但差异不显著。说明常规消毒剂对荧光RT⁃PCR检测结果有影响,特别在对消毒效果评估样品检测时需引起注意。

The emerging novel H5N8 subtype of avian influenza is highly pathogenic and exerts a major threat to the poultry industry. Strengthening monitoring is an important means to prevent and control avian influenza. With the improvement of the biosafety level in the industry,disinfectants have been widely used in various production procedures. How these substances affect the detection results of the current common fluorescent RT-PCR method is rarely reported. In this paper,simulated samples were used to study the effects of disinfectants(i.e.,quicklime,caustic soda,sodium dichloroisocyanate,and glutaraldehyde),detergents(i.e.,washing powder and detergent),protectants(i.e.,glycerol and cephalosporin),and environmental pollutants on the detection results of fluorescent RTPCR. Compared with the corresponding positive control samples,disinfectants had a very significant impact on the detection results. Samples treated with 0.5% quicklime and 0.25% caustic soda could not be detected as positive.Sodium dichloroisocyanate of conventional concentration increased CT value by 10.29 to 10.61,and glutaraldehyde increased CT value by 4.77 to 12.68. Neutralization measures for samples containing alkaline disinfectants could improve the test results. Other detergents,sample protectants and dirt had certain impacts on CT value,but the differences were not significant. The results shows that conventional disinfectants have an impact on the detection results of fluorescent RT-PCR. Attention should be paid especially in the evaluation of disinfection effect.