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Antimicrobial Agents and Chemotherapy, July 2000, p. 1860-1864, Vol. 44, No. 7
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rapid Discriminatory Detection of Genes Coding for SHV beta -Lactamases by Ligase Chain Reaction

Jungmin Kim1,* and Hoan-Jong Lee2

Department of Microbiology, College of Medicine, Dankook University, Cheonan,1 and Department of Pediatrics, Seoul National University College of Medicine, Seoul,2 Korea

Received 11 October 1999/Returned for modification 15 February 2000/Accepted 21 April 2000

Ligase chain reaction (LCR) is a recently developed technique that employs a thermostable ligase and allows for the discrimination of DNA sequences differing in only a single base pair. The method has been adapted and applied to differentiation of blaSHV genes. We have developed an LCR typing method to characterize point mutations in genes for SHV-derived extended-spectrum beta -lactamases with four different sets of biotinylated LCR primers. To evaluate the applicability of the current technique, we tested seven Escherichia coli strains producing SHV-1, SHV-2, SHV-2a, SHV-3, SHV-4, SHV-5, and SHV-12. With the LCR typing, seven SHV genes can be distinguished according to their incorporating point mutations. In an attempt to characterize SHV beta -lactamases by LCR typing in clinical isolates, 46 strains carrying blaSHV genes (32 Klebsiella pneumoniae, 10 Enterobacter cloacae, and 4 E. coli) were subjected to antibiotic susceptibility testing, isoelectric focusing, and LCR typing. LCR typing allowed the characterization of beta -lactamases, and genotypes obtained by LCR typing were in accordance with phenotypes such as antibiotic resistance profile and pI value of beta -lactamase. Therefore, we concluded that LCR typing may permit defining the SHV families with simplicity and reliability and can be applied to the detailed characterization and molecular epidemiology of SHV-type beta -lactamases.

* Corresponding author. Mailing address: Department of Microbiology, College of Medicine, Dankook University, San 29, Anseo-dong, Cheonan, Choongnam, 330-180, Republic of Korea. Phone: 82-417-550-3853. Fax: 82-417-550-3905. E-mail: minkim{at}anseo.dankook.ac.kr.

Antimicrobial Agents and Chemotherapy, July 2000, p. 1860-1864, Vol. 44, No. 7
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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