Open AccessRestriction fragment length dimorphism–PCR method for the detection of extended‐spectrum β‐lactamases unrelated to TEM‐ and SHV‐types
Author(s) -
Lee Sang Hee,
Kim Jae Young,
Shin Sang Heum,
Lee Seok Ki,
Choi Myung Min,
Lee In Yong,
Kim Young Bae,
Cho Jung Youn,
Jin Wook,
Lee Kye Joon
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10708.x
Subject(s) - amplicon , restriction enzyme , biology , polymerase chain reaction , primer (cosmetics) , gene , microbiology and biotechnology , genetics , restriction fragment length polymorphism , in silico pcr , restriction fragment , multiplex polymerase chain reaction , chemistry , organic chemistry
The diagnostic ability of the restriction fragment length dimorphism–polymerase chain reaction (RFLD–PCR) method was evaluated. Seven primer pairs, newly designed from 44 β‐lactamase genes encoding extended‐spectrum β‐lactamases not related to TEM‐ and SHV‐types, were used to differentiate OXA‐2, FOX‐3, CMY‐3, IMP‐1, and IMI‐1 β‐lactamases. The RFLD–PCR was carried out successfully, and these genes were differentiated by the sizes of their PCR products and by the difference in restriction fragment length when each amplicon was digested with a unique restriction enzyme. This discriminatory detection of the genes was confirmed by sequencing the PCR products.