NOVEL PCR ASSAYS FOR THREE FUR MITES IN NATURALLY INFECTED MICE WITHOUT ANIMAL SACRIFICE

Chih-Lin Chou, Ying-Chien Cheng, Ming-Hseng Wang, C. Wan
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Abstract

Rodent fur mite infestation is a persistent and intractable problem in laboratory rodent colonies, due to insensitive diagnostics, unrepresentative samples for testing and improper sentinel system. Myocoptes musculinus (COP), Myobia musculi (MOB) and Radfordia affinis (RDA) have been reported in laboratory mouse colonies. To improve the sensitivity and efficiency of fur mite detection, COP-specific PCR and MOB/RDA-specific PCR assays were developed to detect and differentiate these fur mites, with the existence of a rodent housekeeping gene. The COP-specific PCR and the MOB/RDA-specific PCR could specifically detect COP and MOB/RDA at as low as 10 copies, respectively. In comparison of the specific PCRs with traditional methods (pluck test, tape test and pelt exam) for fur mite diagnosis, 31 rodents and 17 cage environment samples were evaluated. In screening for the infestation status of various fur mites on individual animals, the specific PCR assays showed distinctly higher sensitivity and accuracy (100% and 100%) than those of the traditional methods (sensitivity: 25–80%, accuracy: 83.9–96.8%). Interestingly, by using cage wipe environmental samples, the specific PCR assays exhibited 100% sensitivity and accuracy in the fur mite detection and differentiation. The COP-specific and MOB/RDA-specific PCR assays developed in this study could be reliable alternatives for routine pathogen monitoring of laboratory mice and environments of animal facilities without animal sacrifice. [Chou C-L, Cheng Y-C, Wang MH, Wan CH, Novel PCR assays for three fur mites in naturally infected mice without animal sacrifice, Taiwan Vet J 46(4):133–142, 2020.]
无动物献祭自然感染小鼠三种毛螨的新型PCR检测
由于诊断不敏感、检测样本不具有代表性和哨兵系统不适当,啮齿动物毛螨感染是实验室啮齿动物群体中一个持久而棘手的问题。肌肌病(COP)、肌弱症(MOB)和亲和Radfordia affinis (RDA)在实验室小鼠群体中有报道。为了提高毛螨检测的灵敏度和效率,我们建立了copp特异性PCR和MOB/ rda特异性PCR检测方法来检测和区分毛螨,其中存在啮齿动物内务基因。COP特异性PCR和MOB/RDA特异性PCR分别在低至10拷贝的水平上特异检测到COP和MOB/RDA。对31只啮齿动物和17个鼠笼环境样本进行了特异性pcr检测,并与传统方法(拔毛法、胶带法和毛皮法)进行了比较。在筛选不同动物皮毛螨侵染情况时,特异PCR检测的灵敏度和准确度(100%和100%)明显高于传统方法(灵敏度25 ~ 80%,准确度83.9 ~ 96.8%)。有趣的是,使用笼内擦拭环境样品,特异PCR检测和鉴别皮螨的灵敏度和准确性为100%。本研究建立的cop特异性和MOB/ rda特异性PCR检测方法可作为实验室小鼠和动物设施环境常规病原体监测的可靠替代方法。周春林,程永春,王明明,万超。自然感染小鼠三种毛螨的PCR检测方法[J] .台湾兽医杂志,46(4):133-142,2020。
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