为了解山东地区Kilham细小病毒(kilham rat virus,KRV)的流行情况并探究其分子生物学特征,本试验从某疑似感染KRV的大鼠养殖场分离病毒并对其致病性进行研究。通过观察患病大鼠的临床症状和病理变化并使用PCR方法检测病料确定是否为KRV...为了解山东地区Kilham细小病毒(kilham rat virus,KRV)的流行情况并探究其分子生物学特征,本试验从某疑似感染KRV的大鼠养殖场分离病毒并对其致病性进行研究。通过观察患病大鼠的临床症状和病理变化并使用PCR方法检测病料确定是否为KRV病毒感染。将阳性病料接种到大鼠神经胶质瘤细胞C6上分离KRV并对其VP2全长测序并构建进化树,通过病毒TCID 50测定、血凝和血凝抑制试验及动物回归试验确定分离株的致病性。结果显示养殖场中流产母鼠子宫体积变小,子宫内胎儿全部死亡或部分死亡。将阳性病料接种于C6细胞上,盲传三代后出现明显病变,对分离株VP2基因进行测序发现该分离株与LG/HN/CHN/2016株相似度最高为98.51%,与其他目前已知分离株的VP2同源性在95.20%~97.31%之间。分离株传到7代后的TCID 50为105.56/0.1 mL,血凝效价为1∶1280,血凝抑制效价为27,证明分离株确为KRV,将其命名为KRV/QD/2022。动物回归试验证明该分离株能感染大鼠并导致大鼠出现流产、繁殖障碍等特征性病变。上述结果显示,本研究成功分离出了一株KRV细小病毒并进行了系统的研究,为该病毒的诊断和预防提供了一定的依据。展开更多
Feline panleukopenia virus(FPV)is a single-stranded DNA virus that can infect cats and cause feline panleukopenia,which is a highly contagious and fatal disease in felines.The sequence of FPV is highly variable,and mu...Feline panleukopenia virus(FPV)is a single-stranded DNA virus that can infect cats and cause feline panleukopenia,which is a highly contagious and fatal disease in felines.The sequence of FPV is highly variable,and mutations in the amino acids of its capsid protein play crucial roles in altering viral virulence,immunogenicity,host selection,and other abilities.In this study,the epidemiology of FPV was studied using 746 gastrointestinal swab samples derived from cats that presented gastrointestinal symptoms specifcally,diarrhea or vomiting during the period spanning from 2018 to 2022.The overall prevalence of FPV-positive patients among these samples was determined to be 45.4%.Capsid(virion)protein 2(VP2)gene of each FPV-positive sample was sequenced and amplifed,yielding 65 VP2 sequences.Among them,six VP2 gene sequences were detected in the majority of the samples test positive for FPV,and these positive samples originated from a diverse range of geographical locations.These isolates were named FPV-6,FPV-10,FPV-15,FPV-251,FPV-271 and FPV-S2.Additionally,the substitution of Ala300Pro(A300P)in VP2 was detected for the frst time in feline-derived FPV(FPV-251).FPV-251 isolate,with this substitution in VP2 protein,exhibited stable proliferative capacity in Madin-Darby canine kidney(MDCK)cells and A72 cells.FPV-271 was selected as the FPV control isolate due to its single amino acid diference from VP2 protein of FPV-251 at position 300(FPV-271 has alanine,while FPV-251 has proline).After oral infection,both FPV-251 and FPV-271 isolates caused feline panleukopenia,which is characterized by clinical signs of enterocolitis.However,FPV-251 can infect dogs through the oral route and cause gastrointestinal(GI)symptoms with lesions in the intestine and mesenteric lymph nodes(MLNs)of infected dogs.This is the frst report on the presence of an A300P substitution in VP2 protein of feline-derived FPV.Additionally,FPV isolate with a substitution of A300P at VP2 protein demonstrated efcient replication capabilities in canine cell lines and the ability to infect dogs.展开更多
基金the Experimental Animal Research Project of Hubei Province(Grant No.2023CFA005).
文摘Feline panleukopenia virus(FPV)is a single-stranded DNA virus that can infect cats and cause feline panleukopenia,which is a highly contagious and fatal disease in felines.The sequence of FPV is highly variable,and mutations in the amino acids of its capsid protein play crucial roles in altering viral virulence,immunogenicity,host selection,and other abilities.In this study,the epidemiology of FPV was studied using 746 gastrointestinal swab samples derived from cats that presented gastrointestinal symptoms specifcally,diarrhea or vomiting during the period spanning from 2018 to 2022.The overall prevalence of FPV-positive patients among these samples was determined to be 45.4%.Capsid(virion)protein 2(VP2)gene of each FPV-positive sample was sequenced and amplifed,yielding 65 VP2 sequences.Among them,six VP2 gene sequences were detected in the majority of the samples test positive for FPV,and these positive samples originated from a diverse range of geographical locations.These isolates were named FPV-6,FPV-10,FPV-15,FPV-251,FPV-271 and FPV-S2.Additionally,the substitution of Ala300Pro(A300P)in VP2 was detected for the frst time in feline-derived FPV(FPV-251).FPV-251 isolate,with this substitution in VP2 protein,exhibited stable proliferative capacity in Madin-Darby canine kidney(MDCK)cells and A72 cells.FPV-271 was selected as the FPV control isolate due to its single amino acid diference from VP2 protein of FPV-251 at position 300(FPV-271 has alanine,while FPV-251 has proline).After oral infection,both FPV-251 and FPV-271 isolates caused feline panleukopenia,which is characterized by clinical signs of enterocolitis.However,FPV-251 can infect dogs through the oral route and cause gastrointestinal(GI)symptoms with lesions in the intestine and mesenteric lymph nodes(MLNs)of infected dogs.This is the frst report on the presence of an A300P substitution in VP2 protein of feline-derived FPV.Additionally,FPV isolate with a substitution of A300P at VP2 protein demonstrated efcient replication capabilities in canine cell lines and the ability to infect dogs.
