Sabouraud A, Smith J S, Orciari L A, de Mattos C, de Mattos C, Rohde R
MS G-33 Rabies Laboratory, Viral & Rickettsial Zoonoses Branch, Division of Viral & Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
J Clin Virol. 1999 Jan;12(1):9-19. doi: 10.1016/s1386-6532(98)00006-7.
Alternatives to antigenic typing are needed for epidemiologic surveys of the rabies virus associated with translocated coyotes and foxes, especially in areas where a closely related rabies virus is transmitted by striped skunks.
We developed and evaluated two enzyme based typing methods for rabies virus. The products of a reverse transcription-polymerase chain reaction (RT/PCR) of the nucleoprotein gene were hybridized to type specific probes and detected by enzyme assay after immobilization on microtiter plates.
We tested RT/PCR products of 27 rabies isolates by two different DNA enzyme immunoassays (DEIA) and evaluated the quality of the results from the corresponding nucleotide sequence of the samples.
Using a set of two probes, one of the DEIAs correctly identified 26/27 samples as variants of rabies virus associated with either skunks, foxes, or coyotes. The identity of one fox rabies sample was unresolved by this assay. The second DEIA correctly identified 24/27 samples as variants of rabies virus associated with either skunks, foxes, or coyotes. This assay did not resolve the identity of two fox rabies samples, and misidentified one fox rabies sample as a skunk rabies sample.
DEIA can be used for epidemiologic studies of variants of rabies virus associated with skunks, foxes, and coyotes. Both DEIA methods were effective when typing probes recognized changes at a minimum of two nucleotide positions between variants, but only one assay method was sufficiently stringent to detect a single base pair mismatch. The inherent mutability of RNA viruses must be considered when designing and evaluating typing methods.
对于与迁移的郊狼和狐狸相关的狂犬病病毒进行流行病学调查，需要抗原分型的替代方法，尤其是在条纹臭鼬传播密切相关的狂犬病病毒的地区。
我们开发并评估了两种基于酶的狂犬病病毒分型方法。核蛋白基因的逆转录-聚合酶链反应（RT/PCR）产物与型特异性探针杂交，并在固定于微量滴定板后通过酶测定法进行检测。
我们通过两种不同的DNA酶免疫测定法（DEIA）测试了27株狂犬病病毒分离株的RT/PCR产物，并根据样本的相应核苷酸序列评估了结果的质量。
使用一组两种探针，其中一种DEIA正确地将26/27个样本鉴定为与臭鼬、狐狸或郊狼相关的狂犬病病毒变体。该测定法未确定一个狐狸狂犬病样本的身份。第二种DEIA正确地将24/27个样本鉴定为与臭鼬、狐狸或郊狼相关的狂犬病病毒变体。该测定法未确定两个狐狸狂犬病样本的身份，并将一个狐狸狂犬病样本错误鉴定为臭鼬狂犬病样本。
DEIA可用于与臭鼬、狐狸和郊狼相关的狂犬病病毒变体的流行病学研究。当分型探针识别变体之间至少两个核苷酸位置的变化时，两种DEIA方法均有效，但只有一种测定方法足够严格以检测单个碱基对错配。在设计和评估分型方法时，必须考虑RNA病毒固有的变异性。
