吉隆坡野生大鼠中致病性钩端螺旋体属LipL32基因的培养分离及PCR鉴定
Isolation by culture and PCR identification of LipL32 gene of pathogenic Leptospira spp. in wild rats of Kuala Lumpur.
作者信息
Latifah I, Abdul Halim A, Rahmat M S, Nadia M F, Ubil Z E, Asmah H, Shafariatul Akmar I, Picardeau M, Siti Haslina O, Nasir M A
机构信息
Institute for Medical Research, Jalan Pahang, 50588, Kuala Lumpur, Malaysia.
出版信息
Malays J Pathol. 2017 Aug;39(2):161-166.
BACKGROUND
A study was conducted to confirm the status of rats as the carrier of pathogenic leptospira in Kuala Lumpur, Malaysia.
METHOD
A total of 140 urine samples were collected from trapped rats. These samples were cultured in EMJH enriched media and 18 of these samples (12.9%) were found to be positive when observed under x40 by dark field microscope. Genomic DNA was extracted from all the 18 native isolates for PCR.
RESULT
All the 18 isolates generated the expected 786 base pair band when the set of primers known to amplify LipL32 gene were utilized. These results showed that the primers were suitable to be used for the identification of pathogenic leptospira from the 18 rat samples.
CONCLUSION
The sequencing of the PCR products and BLAST analysis performed on each representative isolates confirmed the pathogenic status of all these native isolates as the LipL32 gene was detected in all the Leptospira isolates. This indicates that the rats are carriers of the pathogenic leptospira in the study area, and therefore are of public health importance.
背景
开展了一项研究以确认马来西亚吉隆坡大鼠作为致病性钩端螺旋体携带者的状况。
方法
从捕获的大鼠中总共收集了140份尿液样本。这些样本在EMJH富集培养基中培养，在暗视野显微镜下40倍观察时，其中18份样本（12.9%）呈阳性。从所有18株本地分离株中提取基因组DNA用于PCR。
结果
当使用已知能扩增LipL32基因的引物组时，所有18株分离株均产生了预期的786碱基对条带。这些结果表明，这些引物适用于从18份大鼠样本中鉴定致病性钩端螺旋体。
结论
对每个代表性分离株进行的PCR产物测序和BLAST分析证实，所有这些本地分离株均具有致病性，因为在所有钩端螺旋体分离株中均检测到了LipL32基因。这表明在研究区域大鼠是致病性钩端螺旋体的携带者，因此具有公共卫生重要性。
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