An rRT-PCR Assay to Detect the Matrix Gene of a Broad Range of Avian Paramyxovirus Serotype-1 Strains

作者：Hines Nichole L; Killian Mary Lea; Pedersen Janice C; Reising Monica M; Mosos Nestor A; Mathieu Benson Christian; Miller Cathy L^*

来源：Avian Diseases, 2012, 56(2): 387-395.

DOI：10.1637/10035-120811-Reg.1

• avian paramyxovirus serotype-1
• Newcastle disease virus
• rRT-PCR
• matrix gene
• pigeon paramyxovirus-1
• National Veterinary Services Laboratories
• National Animal Health Laboratory Network

The current U.S. Department of Agriculture (USDA) validated real-time reverse transcription polymerase chain reaction (rRT-PCR) assay designed to detect the matrix gene of avian paramyxovirus serotype-1 (APMV-1) is the primary screening assay used in the United States. It has previously been shown to be unable to consistently detect all members of class I APMV-1. Diagnostic testing relies on rRT-PCR to quickly detect APMV-1 in wild birds, backyard flocks, live bird markets, commercial poultry, and for export testing. Limitations of the current USDA assay have raised concerns about the potential for some strains of APMV-1 to remain undetected by the primary screening assay. Mismatches in the probe were shown to cause a loss in template binding efficiency, resulting in lack of detection by the assay. Here, we describe the development and analytical validation of a new rRT-PCR assay designed to target a highly conserved region of the matrix gene across a wide range of APMV-1 strains. Limit of detection testing revealed a 3 log(10) decrease in sensitivity for one low-virulence strain when compared to the USDA validated assay. Conversely, the assay showed increased sensitivity for a class I isolate and two virulent strains of APMV-1 that were not detected by the USDA-validated assay. The new assay also demonstrated a high degree of specificity by the lack of detection of 43 non APMV-1 viruses.

• 出版日期2012-6