Smreczak Marcin, Orłowska Anna, Müller Thomas, Freuling Conrad M, Kawiak-Sadurska Magdalena, Trębas Paweł
Department of Virology, National Veterinary Research Institute, 24-100 Puławy, Poland.
Institute for Molecular Virology and Cell Biology, Friedrich-Loeffler-Institute (FLI), 17493 Greifswald-Insel Riems, Germany.
J Vet Res. 2022 Nov 15;66(4):473-477. doi: 10.2478/jvetres-2022-0065. eCollection 2022 Dec.
Rabies as a zoonosis threatens public health worldwide. Several thousand people die each year of infections by the rabies virus (RABV). Oral rabies vaccination (ORV) of wildlife was successfully implemented in many European countries and led to rabies being brought under control there. In Poland, ORV was introduced in 1993 using vaccines containing an attenuated strain of the rabies virus. However, attenuated rabies viruses may have residual pathogenicity and cause the disease in target and non-target animals.
A red fox carcass was tested as part of national rabies surveillance, and its brain was screened for RABV infection using two conjugates and a fluorescent antibody test (FAT). The rabies virus was isolated in mouse neuroblastoma cells by rabies tissue culture infection test (RTCIT), and viral RNA was detected by heminested reverse transcriptase PCR (hnRT-PCR) as well as by quantitative real-time RT-PCR (rtRT-qPCR). An amplicon of 600 bp was subjected to Sanger sequencing. To differentiate between vaccine and field RABV strains, PCR-restriction fragment length polymorphism (PCR-RFLP) using the Dra I, Msp I, Nla IV and Mbo II restriction endonucleases was performed.
The rabies virus was detected in the fox's brain using FAT, RTCIT and molecular tests. The PCR-RFLP revealed of vaccine-induced rabies, and full-length genome analysis showed 100% nucleotide sequence identity of the isolate with the reference sequences of Street Alabama Dufferin Bern (SAD Bern) vaccine strains and other vaccine-induced rabies virus isolates detected in animals and deposited in GenBank.
We detected vaccine-induced rabies for the first time in Poland in a fox during routine rabies surveillance.
狂犬病作为一种人畜共患病,威胁着全球公共卫生。每年有数千人死于狂犬病病毒(RABV)感染。野生动物口服狂犬病疫苗(ORV)在许多欧洲国家成功实施,并使狂犬病在当地得到控制。在波兰,1993年开始使用含有减毒狂犬病病毒株的疫苗进行ORV。然而,减毒狂犬病病毒可能具有残余致病性,并在目标动物和非目标动物中引发疾病。
作为国家狂犬病监测的一部分,对一只赤狐尸体进行了检测,并使用两种结合物和荧光抗体试验(FAT)对其大脑进行RABV感染筛查。通过狂犬病组织培养感染试验(RTCIT)在小鼠神经母细胞瘤细胞中分离出狂犬病病毒,并通过半巢式逆转录聚合酶链反应(hnRT-PCR)以及定量实时逆转录聚合酶链反应(rtRT-qPCR)检测病毒RNA。对一个600 bp的扩增子进行桑格测序。为区分疫苗株和野毒株RABV,使用Dra I、Msp I、Nla IV和Mbo II限制性内切酶进行了聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)。
通过FAT、RTCIT和分子检测在狐狸大脑中检测到狂犬病病毒。PCR-RFLP显示为疫苗诱导的狂犬病,全长基因组分析表明,分离株与阿拉巴马街达弗林伯恩(SAD Bern)疫苗株的参考序列以及在动物中检测到并保存在GenBank中的其他疫苗诱导的狂犬病病毒分离株的核苷酸序列同一性为100%。
在波兰的常规狂犬病监测中,我们首次在一只狐狸中检测到疫苗诱导的狂犬病。