摘要
【目的】建立禽偏肺病毒乳胶凝集试验检测方法,以快速、有效、准确地检测该病毒。【方法】用制备的禽偏肺病毒抗体致敏乳胶,与禽偏肺病毒反应,优化最佳致敏条件,建立禽偏肺病毒乳胶凝集试验检测方法;对该方法的重复性与特异性进行检测,将该方法与套式RT-PCR进行比较,并将其用于临床病料的检测。【结果】禽偏肺病毒抗体致敏乳胶的最佳致敏温度为56℃,抗体最佳稀释度为1∶10(体积比),最佳致敏时间为120min。用建立的乳胶凝集试验检测方法和套式RT-PCR同步检测10份禽偏肺病毒尿囊液,两者的阳性检出率一致,总符合率为100%。应用该方法对采自山东省不同地区的68份可疑临床病料进行检测,阳性检出率为26.47%。【结论】建立了禽偏肺病毒乳胶凝集试验检测方法,该法具有快速、简便、准确、特异性强、重复性和稳定性良好等优点,是一种适于基层单位检测禽偏肺病毒的可靠方法。
[Objective] This study aimed to establish latex agglutination test （LAT） for detecting Avi- an metapneumovirus （aMPV） rapidly,effectively and accurately. [Method] LAT was established based on suitable conditions for sensitizing the latex determined by titration test. Repeatability and specificity were assessed compared with nested RT PCR, and clinical samples were collected and tested by LAT as well. [Result] The best reaction conditions were： antibody dilution 1 ： 10, reaction temperature 56 ~C and reac- tion time 2 hours. 10 samples of aMPV allantoic fluid were tested by LAT and nested RT-PCR,the positive rates were identical and the total coincidence rate was 100%. 68 samples suspected of infection with aMPV, were detected by LAT and the positive rate was 26.47%. [Conclusion] The established LAT method with advantages of rapidity, simplicity, high accuracy, specificity and also repeatability and stability can be used for clinical diagnosis of aMPV infection.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2014年第2期24-28,共5页
Journal of Northwest A&F University(Natural Science Edition)
基金
山东省科技发展计划项目(2010GNC10912)
关键词
禽偏肺病毒
乳胶凝集试验
检测
Avian metapneumovirus
latex agglutination test （LAT）
detection
