Rapid Identification of Avian Mycoplasma Using Nested and Species-Specific Polymerase Chain Reaction

以巢式及種別特異聚合酶連鎖反應快速鑑定家禽黴漿菌

劉宏仁(Hung-Jen Liu) ； 柯冠銘(Guan-Ming Ke) ； 徐筱微(Hsiao-Wei Hsu) ； 林茂勇(Maw-Yeong Lin) ； 廖明輝(Ming-Huei Liao) ； 吳永惠(Yeong-Huey Wu)

《中華民國獸醫學會雜誌》 27卷1期 (2001/03) Pp. 1-8

https://doi.org/10.7009/JTCSVS.200103.0001

摘要

家禽黴漿菌感染常造成家禽產業之經濟損失而為業界所關切。為能快速及正確診斷家禽黴漿菌感染症，因此建立種別特異及巢式聚合酶連鎖反應快速鑑定至少九種標準家禽黴漿菌菌種及野外家禽黴漿菌感染病例。為確認聚合酶連鎖反應之特異性，擴增之產物經限制酶分析確認。在敏感度試驗，巢式聚合酶連鎖反應至少可提高聚合酶連鎖反應之敏感度達1,000倍。因此，巢式聚合酶連鎖反應適於偵測早期感染或病變組織內極微量之家禽黴漿菌。若配合種別特異(species-specific)之引子可進一步鑑定家禽黴漿菌種。本研究結果顯示種別特異及巢式聚合酶連鎖反應可快速、特異及敏感地偵測標準家禽黴漿菌菌株及並可應用於鑑定野外家禽黴漿菌感染症。

關鍵字

家禽黴漿菌 ； 限制酶 ； 種別特異及巢式聚合酶連鎖反應

並列摘要

Mycoplasma infections are of concern in avian medicine, because they cause economic losses in commercial poultry production. A nested polymerase chain reaction (PCR) was established to detect nine reference species and field isolates of avian mycoplasma. To confirm the specificity of PCR, PCR products were digested with restriction enzymes. Field isolates confirmed to be avian mycoplasma were further identified by using a species-specific PCR assay. The results showed that nested PCR was able to increase the PCR sensitivity by approximately 103 folds. Amplification with the nested primers allows to detect early in the infection when the quantity of avian mycoplasmas in the tissue is very low. Our results suggested that the species-specific and nested PCR assays were a rapid, sensitive, and specific approach to identify each species of avian mycoplasmas.

並列關鍵字

Avian mycoplasma ； Restriction enzyme ； Species-specific and nested polymerase chain reaction