Hussain I, Wani S A, Qureshi S D, Farooq S
Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shuhama (Alusteng), Srinagar 190006, India.
Mol Cell Probes. 2009 Apr;23(2):112-4. doi: 10.1016/j.mcp.2009.01.003. Epub 2009 Jan 21.
One hundred and twenty-eight swab samples from footrot lesions of naturally infected sheep were examined for presence of Dichelobacter nodosus (D. nodosus). The detection of D. nodosus was carried out by polymerase chain reaction (PCR), directly from swabs or after isolation, using 16S rDNA specific primers. The isolation of the bacterium was carried out anaerobically on trypticase-arginine-serine (TAS) agar containing 4% hoof powder. Serogrouping of the D. nodosus was accomplished with multiplex PCR using nine (A-I) serogroup specific primers. The virulent and benign status of the isolates was ascertained by detection of virulence specific integrase A (intA) gene. Out of total 83 D. nodosus isolates, 62 (74.69%) belonged to serogroup B, 18 (21.68%) to serogroup E and three (3.62%) to serogroup I. Serogroup I was detected and isolated for the first time in India. All the positive samples revealed infection by single serogroup of D. nodosus except one which showed mixed infection of serogroups B and E. Sixty (72.28%) isolates possessed intA gene and thus were considered as virulent strains. Serogroupwise intA gene was found in 43 (69.35%) isolates of serogroup B, 14 (77.78%) of E and in all the three (100%) of I.
对128份来自自然感染绵羊蹄腐病病变部位的拭子样本进行检测，以确定是否存在坏死梭杆菌（D. nodosus）。通过聚合酶链反应（PCR）直接从拭子或分离培养后使用16S rDNA特异性引物检测坏死梭杆菌。该细菌在含有4%蹄粉的胰蛋白酶-精氨酸-丝氨酸（TAS）琼脂上进行厌氧分离培养。使用9种（A-I）血清群特异性引物通过多重PCR对坏死梭杆菌进行血清群分类。通过检测毒力特异性整合酶A（intA）基因确定分离株的毒力和良性状态。在总共83株坏死梭杆菌分离株中，62株（74.69%）属于血清群B，18株（21.68%）属于血清群E，3株（3.62%）属于血清群I。血清群I在印度首次被检测到并分离出来。除1份样本显示血清群B和E混合感染外，所有阳性样本均显示由单一血清群的坏死梭杆菌感染。60株（72.28%）分离株具有intA基因，因此被认为是毒力菌株。血清群B的43株（69.35%）分离株、血清群E的14株（77.78%）分离株以及血清群I的所有3株（100%）分离株均发现有intA基因。
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