Open AccessCharacterisation of extended‐spectrum β‐lactamases of the SHV family using a combination of PCR‐single strand conformational polymorphism (PCR‐SSCP) and PCR‐restriction fragment length polymorphism (PCR‐RFLP)
Author(s) -
Chanawong Aroonwadee,
M'Zali Fatima H.,
Heritage John,
Lulitad Aroonlug,
Hawkey Peter M.
Publication year - 2000
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.1574-6968.2000.tb08995.x
Subject(s) - single strand conformation polymorphism , restriction fragment length polymorphism , biology , polymerase chain reaction , genetics , restriction enzyme , microbiology and biotechnology , gene
Polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) has been developed to extend the identification of SHV β‐lactamases previously characterised by PCR‐single strand conformational polymorphism (PCR‐SSCP) analysis alone. Eight bacteria, each producing a different SHV β‐lactamase, were used in this study. These bacteria harbour bla SHV‐1 , bla SHV‐2a , bla SHV‐3 , bla SHV‐4 , bla SHV‐5 (two strains), bla SHV‐11 and bla SHV‐12 . All isolates were characterised by PCR‐SSCP and PCR‐RFLP with Dde I and Nhe I digestion. By a combination of these techniques, the genes encoding these β‐lactamases could be differentiated from each other. In addition, the PCR‐RFLP technique theoretically can be applied to distinguish the genes encoding SHV‐7, SHV‐9, SHV‐10, SHV‐15, SHV‐17 and SHV‐24 from those encoding other SHV variants. We report a simple PCR‐RFLP technique that can be used in epidemiological studies to enable the rapid characterisation of known SHV β‐lactamases in a combination with the previously published PCR‐SSCP analysis.