Development of multiplex polymerase chain reaction for the detection of vancomycin resistant genotypes and Enterococcus Sp.-specific genes

Cho, Yun-Sang; Lee, Hee-Soo; Kim, Jong-Man; Ahn, Jong-Sam; Ryu, Pan-Dong; Park, Yong-Ho; Yoo, Han-Sang; Lee, Mun-Han

Korean Journal of Veterinary Research 2003;43(1):103-112.

Yun-Sang Cho¹, Hee-Soo Lee¹, Jong-Man Kim¹, Jong-Sam Ahn¹, Pan-Dong Ryu², Yong-Ho Park², Han-Sang Yoo², Mun-Han Lee²

¹National Veterinary Research and Quarantine Service
²College of Veterinary Medicine, Seoul National University

장구균의 vancomycin 내성 유전자와 종 특이유전자의 검출을 위한 Multiplex polymerase chain reaction 개발

조윤상¹, 이희수¹, 김종만¹, 안종삼¹, 류판동², 박용호², 유한상², 이문한²

¹국립수의과학검역원
²서울대학교 수의과대학

Abstract

A multiplex PCR assay, which allows simultaneous detection of vancomycin resistant genotypes and Enterococcus species-specific genes, was developed. Vancomycin resistant enterococci (VRE) from chickens and humans could be detected for vanA, vanB, vanC-1, vanC-2, $ddl_{E.faecium}$ and $ddl_{E.faecalis}$ by multiplex PCR. Eight isolates of VRE from humans (n=11) had $ddl_{E.faecium}$ and vanA, and 3 isolates of the VRE had $ddl_{E.faecium}$ and vanB. One isolate of VRE from chickens (n=6) had $ddl_{E.faecium}$ and vanA, and 5 isolates of the VRE had only vanA. E. faecium, E. faecalis, E. gallinarum and E. casseliflavus were also confirmed for the species-specific gene by multiplex PCR. This multiplex PCR could detect E. faecium, E. faecalis, E. gallinarum, E. casseliflavus, vanA, vanB, vanC-1 and vanC-2, simultaneously. The PCR assay established in the present study can be an alternative to time-consuming biochemical tests and antibiotic susceptibility tests of Enterococcus spp.

Key Words: vancomycin resistant enterococci, multiplex polymerase chain reaction