Trinh T T Hang, Do V Tan, Do V Khien, Vu-Khac Hung
Department of Biotechnology, Institute of Veterinary Research and Development of Central Vietnam, Nha Trang City, Vietnam.
Department of Virology, Institute of Veterinary Research and Development of Central Vietnam, Nha Trang City, Vietnam.
Vet World. 2024 Jul;17(7):1530-1537. doi: 10.14202/vetworld.2024.1530-1537. Epub 2024 Jul 13.
No study has successfully isolated parvovirus in Vietnam. This study aimed to isolate and characterize parvovirus strains indigenous in Vietnam for vaccine development against porcine parvovirus (PPV).
We collected serum and stillbirth samples from six provinces in Vietnam, and PPV-positive samples were identified using a polymerase chain reaction. Parvovirus isolation was attempted using the PK-15 cells maintained in a minimum essential medium supplemented with 5% fetal bovine serum and 1% antibiotics (Penicillin-streptomycin). The cells were incubated at 37°C with 5% CO. Virulence experiments were conducted on white primiparous sows to evaluate the virulence of the PPV strain through hemagglutination inhibition (HI) titers and fetus lesions.
We analyzed 360 serum and 32 stillbirth (liver and lungs) samples, revealing that 32/392 (8.2% ) of them were PPV-positive, all belonging to PPV1. Thirty-two PPV-positive samples were successfully isolated, with 100% identity as VP2 sequences. The phylogenetic tree revealed a close relationship with the Kresse strain (isolated from Canada in 1996) and the PPV1-0225-L-SD strain (isolated from China in 2022). Two PPV isolates (VC5 from Dongnai and TX7 from Thanhhoa) that exhibited high 50% tissue culture infectious dose titers were selected for the virulence experiment. On day 21, after injection, the HI antibody titers ranged from 10log to 12log. On day 90, 71%-80% of fetuses were mummified.
This study showed that the PPV infection rate in Vietnam was 8.2%. Thirty-two isolates belonged to PPV1. Two PPV strains, VC5 and TX7, were determined to be highly virulent by the results of HI titers after injection into gilts. VC5 and TX7 were determined to be good candidates for further research on PPV vaccines.
越南尚未有研究成功分离出细小病毒。本研究旨在分离并鉴定越南本土的细小病毒株,用于开发抗猪细小病毒(PPV)疫苗。
我们从越南六个省份收集了血清和死胎样本,并使用聚合酶链反应鉴定PPV阳性样本。尝试使用在添加5%胎牛血清和1%抗生素(青霉素-链霉素)的最低必需培养基中培养的PK-15细胞分离细小病毒。细胞在37°C、5%二氧化碳条件下孵育。对白色初产母猪进行毒力实验,通过血凝抑制(HI)效价和胎儿病变评估PPV毒株的毒力。
我们分析了360份血清样本和32份死胎(肝脏和肺)样本,发现其中32/392(8.2%)为PPV阳性,均属于PPV1。成功分离出32份PPV阳性样本,其VP2序列的同源性为100%。系统发育树显示与Kresse毒株(1996年从加拿大分离)和PPV1-0225-L-SD毒株(2022年从中国分离)关系密切。选择两个表现出高50%组织培养感染剂量效价的PPV分离株(同奈省的VC5和清化省的TX7)进行毒力实验。注射后第21天,HI抗体效价范围为10log至12log。第90天,71%-80%的胎儿木乃伊化。
本研究表明越南PPV感染率为8.2%。32个分离株属于PPV1。注射后备母猪后,根据HI效价结果确定VC5和TX7这两个PPV毒株具有高毒力。VC5和TX7被确定为PPV疫苗进一步研究的良好候选毒株。