Dhanashree B, Otta S K, Karunasagar I, Karunasagar I
Department of Microbiology, Kasturba Medical College, Mangalore, India.
Indian J Med Res. 2003 Jan;117:19-24.
BACKGROUND & OBJECTIVES: Listeria monocytogenes is an important food-borne pathogen causing meningitis and septicaemia in newborns and immunocompromised persons, abortion and preterm labour in pregnant women. Though various methods are available for typing L. monocytogenes, RAPD analysis has been used for epidemiological purposes in developed countries due to its greater discriminating ability. However, as there are no published reports from India on the typing of L. monocytogenes by RAPD technique the present study was undertaken to type isolates of L. monocytogenes from clinical, food and veterinary samples.
Isolates of L. monocytogenes were subjected to RAPD using four decamer random primers R1, R2, R3 and R4. Amplified products were analysed by agarose gel electrophoresis.
Eight strains of L. monocytogenes on RAPD analysis generated 4 distinct profiles each with R1 and R4 primers and 3 different profiles with R2 and R3 primers. The isolates from fish, clinical and veterinary samples showed different profiles with respect to each other. Isolate from flat fish (serovar 4) showed a different profile from that of clams (serovar 1). Two isolates from placenta (serovar 1) showed similar profiles and all the isolates from veterinary samples generated similar profiles.
INTERPRETATION & CONCLUSION: RAPD analysis in the present study allowed discrimination of isolates among the same serotype but from different sources. Since RAPD is a rapid technique and offers greater discrimination of strains, this method may be used for typing L. monocytogenes in India.
单核细胞增生李斯特菌是一种重要的食源性病原体,可导致新生儿和免疫功能低下者患脑膜炎和败血症,使孕妇流产和早产。尽管有多种方法可用于单核细胞增生李斯特菌的分型,但由于随机扩增多态性DNA(RAPD)分析具有更强的鉴别能力,在发达国家已被用于流行病学研究。然而,由于印度尚无关于用RAPD技术对单核细胞增生李斯特菌进行分型的报道,因此本研究旨在对临床、食品和兽医样本中的单核细胞增生李斯特菌分离株进行分型。
使用四种十聚体随机引物R1、R2、R3和R4对单核细胞增生李斯特菌分离株进行RAPD分析。扩增产物通过琼脂糖凝胶电泳进行分析。
对八株单核细胞增生李斯特菌进行RAPD分析,使用R1和R4引物时各产生4种不同的图谱,使用R2和R3引物时产生3种不同的图谱。来自鱼类、临床和兽医样本的分离株彼此显示出不同的图谱。比目鱼(血清型4)的分离株与蛤(血清型1)的分离株图谱不同。来自胎盘的两株分离株(血清型1)显示出相似的图谱,所有来自兽医样本的分离株产生相似的图谱。
本研究中的RAPD分析能够区分同一血清型但来源不同的分离株。由于RAPD是一种快速技术且对菌株具有更强的鉴别能力,该方法可用于印度单核细胞增生李斯特菌的分型。