Zaher Kawther Sayed, El-Dabae Wahid Hussein, El-Sebelgy Mostafa Mohamed, Aly Naglaa Ibrahim, Salama Zeinab Taha
Department of Microbiology and Immunology, Veterinary Research Division, National Research Centre, Dokki 12622, Giza, Egypt.
Department of Pet Animal Vaccine Research, Veterinary Serum and Vaccine Research Institute, Abbasia, Egypt.
Vet World. 2020 Feb;13(2):326-333. doi: 10.14202/vetworld.2020.326-333. Epub 2020 Feb 19.
This study aimed to detect and characterize current genotypes of canine parvovirus (CPV) in Egypt during 2018.
A total of 50 fecal swabs were collected from clinically infected domestic dogs of 2-5 months of age, suspected to suffer from CPV infection, from Cairo and Giza Governorates. The samples were subjected to qualitative antigen detection using the rapid test, followed by isolation on Madin-Darby Canine Kidney (MDCK) cells, molecular characterization with partial amplification of VP2 gene using polymerase chain reaction (PCR), followed by sequencing and phylogenetic analysis.
Out of 50 fecal samples, 20 samples were positive (40%) by Rapid CPV/canine coronavirus Ag Test Kit. These positive samples were cultured successfully on MDCK cells. Nine randomly chosen samples out of 30 apparently negative samples were amplified using PCR with primers Hfor and Hrev to yield a typical 630 bp fragment. Then, six randomly chosen samples out of nine were amplified using PCR with primers Pbs and Pbas to yield a typical 427 bp fragment. Sequencing, BLAST analysis and assembly of the two fragments (630 bp and 427 bp) to produce 912 bp fragments, in the six samples, revealed two serotypes CPV-2b and CPV-2c. The obtained strains were submitted to GenBank and given accession numbers MK642272, MK642273, MK642274, MK642275, MK642276, and MK642277. Phylogenetic analysis of the Egyptian strains serotype 2b illustrated that they were closely related to Thailand strains (accession numbers KP715709, KP715694, KP715701, and KP715700); while Egyptian strains serotype 2c was closely related to Thailand strains (accession numbers MH711894 and MH711902), Taiwanese strain (KU244254), Chinese strain (MF467242), and Vietnamese strain (accession number LC216910).
The current research recommends further epidemiological studies to assess the extent of the occurrence of different serotypes of CPV in Egypt and the efficiency of imported and locally produced vaccines in protection against CPV infection.
本研究旨在检测并鉴定2018年埃及犬细小病毒（CPV）的当前基因型。
从开罗和吉萨省2至5月龄临床感染疑似患有CPV感染的家犬中总共采集了50份粪便拭子。使用快速检测法对样本进行定性抗原检测，随后在麦迪逊-达比犬肾（MDCK）细胞上进行分离培养，利用聚合酶链反应（PCR）对VP2基因进行部分扩增以进行分子鉴定，随后进行测序和系统发育分析。
在50份粪便样本中，20份样本通过犬细小病毒/犬冠状病毒抗原快速检测试剂盒检测为阳性（40%）。这些阳性样本在MDCK细胞上成功培养。从30份看似阴性的样本中随机选取9份样本，使用引物Hfor和Hrev进行PCR扩增，得到一个典型的630 bp片段。然后，从这9份样本中随机选取6份样本，使用引物Pbs和Pbas进行PCR扩增，得到一个典型的427 bp片段。对这6份样本中两个片段（630 bp和427 bp）进行测序、BLAST分析并组装成912 bp片段，结果显示有两种血清型，即CPV-2b和CPV-2c。所获得的菌株已提交至GenBank，并被赋予登录号MK642272、MK642273、MK642274、MK642275、MK642276和MK642277。对埃及2b血清型菌株的系统发育分析表明，它们与泰国菌株（登录号KP715709、KP715694、KP715701和KP715700）密切相关；而埃及2c血清型菌株与泰国菌株（登录号MH711894和MH711902）、台湾菌株（KU244254）、中国菌株（MF467242）和越南菌株（登录号LC216910）密切相关。
当前研究建议进一步开展流行病学研究，以评估埃及不同血清型CPV的发生程度以及进口和本地生产疫苗预防CPV感染的效果。
