A set of RT-PCR assays for detection of all known avian paramyxoviruses and application in surveillance of avian paramyxoviruses in China
Ji-Hui Jin,
Jing-Jing Wang,
Ying-Chao Ren,
Shuo Liu,
Jin-Ping Li,
Guang-Yu Hou,
Hua-Lei Liu,
Qing-Ye Zhuang,
Su-Chun Wang,
Wen-Ming Jiang,
Xiao-Hui Yu,
Jian-Min Yu,
Li-Ping Yuan,
Cheng Peng,
Guo-Zhong Zhang,
Ji-Ming Chen
Affiliations
Ji-Hui Jin
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Jing-Jing Wang
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Ying-Chao Ren
Department for Animal Health Assessment, China Animal Health and Epidemiology Center, Qingdao, China
Shuo Liu
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Jin-Ping Li
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Guang-Yu Hou
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Hua-Lei Liu
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Qing-Ye Zhuang
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Su-Chun Wang
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Wen-Ming Jiang
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Xiao-Hui Yu
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Jian-Min Yu
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Li-Ping Yuan
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Cheng Peng
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Guo-Zhong Zhang
Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, China
Ji-Ming Chen
Laboratory for Avian Disease Surveillance (OIE Reference Laboratory for Newcastle Disease), China Animal Health and Epidemiology Center, Qingdao, China
Background Avian paramyxoviruses (APMVs), also termed avian avulaviruses, are of a vast diversity and great significance in poultry. Detection of all known APMVs is challenging, and distribution of APMVs have not been well investigated. Methods A set of reverse transcription polymerase chain reaction (RT-PCR) assays for detection of all known APMVs were established using degenerate primers targeting the viral polymerase L gene. The assays were preliminarily evaluated using in-vitro transcribed double-stranded RNA controls and 24 known viruses, and then they were employed to detect 4,346 avian samples collected from 11 provinces. Results The assays could detect 20–200 copies of the double-stranded RNA controls, and detected correctly the 24 known viruses. Of the 4,346 avian samples detected using the assays, 72 samples were found positive. Of the 72 positives, 70 were confirmed through sequencing, indicating the assays were specific for APMVs. The 4,346 samples were also detected using a reported RT-PCR assay, and the results showed this RT-PCR assay was less sensitive than the assays reported here. Of the 70 confirmed positives, 40 were class I Newcastle disease virus (NDV or APMV-1) and 27 were class II NDV from poultry including chickens, ducks, geese, and pigeons, and three were APMV-2 from parrots. The surveillance identified APMV-2 in parrots for the first time, and revealed that prevalence of NDVs in live poultry markets was higher than that in poultry farms. The surveillance also suggested that class I NDVs in chickens could be as prevalent as in ducks, and class II NDVs in ducks could be more prevalent than in chickens, and class II NDVs could be more prevalent than class I NDVs in ducks. Altogether, we developed a set of specific and sensitive RT-PCR assays for detection of all known APMVs, and conducted a large-scale surveillance using the assays which shed novel insights into APMV epidemiology.
