Development of a Molecular Assay for Laboratory Rodent Fur Mite Diagnosis and Comparison with the Traditional Diagnostic Methods
| Autor: | Chih-Lin Chou, 周知林 |
|---|---|
| Rok vydání: | 2019 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 107 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. To improve the sensitivity and efficiency of fur mite detection, a multiplex PCR assay was developed to simultaneously detect and differentiate different species of fur mites, including Myocoptes musculinus (COP), Myobia musculi (MOB) and/or Radfordia spp. (RAD), and species A (SPA; a novel rodent fur mite identified in Taiwan), with the existence of a rodent housekeeping gene. This multiplex PCR could specifically detect as low as 10 copies of each species in equal-amount triple infestation. Super-infestation with 10 to 100-fold differences in mite burdens could be also detected. In comparison of the multiple PCR and traditional methods (pluck test, tape test, and pelt exam) for fur mite diagnosis, 48 rodents and 25 cage environment samples were evaluated for the fur mite infestation. In screening the status of various fur mites on individual animals, the multiplex PCR assay showed distinctly higher in sensitivity and accuracy (86 % and 95.1 %) than that of traditional methods (sensitivity: 6 % - 46 %, accuracy: 67.4 % - 81.3 %). Interestingly, by using cage wipe environmental samples, the multiplex PCR assay exhibited 100 % in both sensitivity and accuracy on the fur mite detection and differentiation. The COP/MOB-RAD/SPA/Actin multiplex PCR assay developed in this study could be a reliable alternative method for routine pathogen monitoring (animal or environment) or for tracing the suspect fur mite outbreak in rodent colonies. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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