Arun Paravalappil Muraleedharan, Rajasekhar Ravindran, Ravishankar Chintu, Palekkodan Hamza, Kanjirakkuzhiyil Sumod, Somasekhar Shashank
Departments of Veterinary Microbiology, College of Veterinary and Animal Sciences, Kerala Veterinary and Animal Sciences University, Pookode, Kerala, India.
Veterinary Pathology, College of Veterinary and Animal Sciences, Kerala Veterinary and Animal Sciences University, Pookode, Kerala, India.
J Vet Diagn Invest. 2025 Mar;37(2):278-283. doi: 10.1177/10406387241311514. Epub 2025 Jan 27.
Peste-des-petits-ruminants (PPR) is primarily a disease of small ruminants caused by peste-des-petits-ruminants virus (PPRV; , ), formerly the small ruminant morbillivirus. PPRV can cause significant morbidity and mortality in small ruminants and a significant economic impact. Conventional reverse-transcription PCR (RT-PCR), and probe-based and SYBR Green-based RT quantitative real-time PCR (RT-qPCR), are employed for the molecular detection of PPRV. Here we describe a SYBR Green-based RT-qPCR for rapid and sensitive detection of PPRV. We designed the specific primers from the conserved region of the fusion gene () of PPRV. The standard curve of the established RT-qPCR assay had a good linear relationship. The developed assay was also 3 log units more sensitive than the conventional RT-PCR, with a detection limit of 13.6 copies and an efficiency of 98.2%. There was no cross-reactivity with other caprine respiratory viruses, namely bluetongue virus, goatpox virus, and orf virus. The positive detection rate of clinical samples was 11 of 64 (17.2%) versus 10 of 64 (15.6%) by conventional RT-PCR. We confirmed our results by sequencing the full and genes of the isolates. Our SYBR Green RT-qPCR can be used as a fast, economical, and sensitive alternative to RT-PCR for the detection of PPRV.
小反刍兽疫（PPR）主要是由小反刍兽疫病毒（PPRV； ， ）引起的小反刍动物疾病，PPRV以前称为小反刍兽麻疹病毒。PPRV可在小反刍动物中导致显著的发病率和死亡率，并造成重大经济影响。传统的逆转录PCR（RT-PCR）以及基于探针和基于SYBR Green的RT定量实时PCR（RT-qPCR）被用于PPRV的分子检测。在此，我们描述一种基于SYBR Green的RT-qPCR方法，用于快速灵敏地检测PPRV。我们从PPRV融合基因（ ）的保守区域设计了特异性引物。所建立的RT-qPCR检测方法的标准曲线具有良好的线性关系。所开发的检测方法比传统RT-PCR灵敏3个对数单位，检测限为13.6个拷贝，效率为98.2%。与其他山羊呼吸道病毒，即蓝舌病毒、山羊痘病毒和口疮病毒无交叉反应。临床样本的阳性检出率通过传统RT-PCR为64份中的10份（15.6%），而通过本方法为64份中的11份（17.2%）。我们通过对分离株的完整 和 基因进行测序证实了结果。我们的SYBR Green RT-qPCR可作为一种快速、经济且灵敏的替代方法，用于替代RT-PCR检测PPRV。
