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Antimicrobial Agents and Chemotherapy, January 2006, p. 204-209, Vol. 50, No. 1
0066-4804/06/$08.00+0     doi:10.1128/AAC.50.1.204-209.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Simultaneous Detection of Nine Antibiotic Resistance-Related Genes in Streptococcus agalactiae Using Multiplex PCR and Reverse Line Blot Hybridization Assay{dagger}

Xianyu Zeng,1,2 Fanrong Kong,1 Hui Wang,1,2 Archie Darbar,1,{ddagger} and Gwendolyn L. Gilbert1*

Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Westmead, New South Wales, Australia,1 Department of Dermatology, Wuhan First Hospital, Wuhan, Hubei Province, People's Republic of China2

Received 24 July 2005/ Returned for modification 28 August 2005/ Accepted 24 October 2005

Streptococcus agalactiae (group B streptococcus [GBS]) is the leading cause of neonatal and maternal sepsis. Penicillin is recommended for intrapartum prophylaxis, but erythromycin or clindamycin is used for penicillin-allergic carriers. Antibiotic resistance (AR) has increased recently and needs to be monitored. We have developed a multiplex PCR-based reverse line blot (mPCR/RLB) hybridization assay to detect, simultaneously, seven genes encoding AR—erm(A/TR), erm(B), mef(A/E), tet(M), tet(O), aphA-3, and aad-6—and two AR-related genes, int-Tn and mreA. We tested 512 GBS isolates from Asia and Australasia and compared mPCR/RLB with antibiotic susceptibility phenotype or single-gene PCR. Phenotypic resistance to tetracycline was identified in 450 (88%) isolates, of which 442 had tet(M) (93%) and/or tet(O) (6%). Of 67 (13%) erythromycin-resistant isolates, 18 were susceptible to clindamycin, i.e., had the M phenotype, encoded by mef(A/E); 39 had constitutive (cMLSB) and 10 inducible clindamycin resistance, and of these, 34 contained erm(B) and 12 erm(A/TR). Of four additional isolates with mef(A/E), three contained erm(B) with cMLSB and one was erythromycin susceptible. Of 61 (12%) clindamycin-resistant isolates, 20 were susceptible to erythromycin and two had intermediate resistance. Based on sequencing, 21 of 22 isolates with mef had mef(E), and 8 of 353 with int-Tn had an atypical sequence. Several AR genes, erm(B), tet(O), aphA-3, aad-6, and mef(A/E), were significantly more common among Asian than Australasian isolates, and there were significant differences in distribution of AR genes between GBS serotypes. Our mPCR/RLB assay is simple, rapid, and suitable for surveillance of antibiotic resistance in GBS.

* Corresponding author. Mailing address: Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, Darcy Road, Westmead, New South Wales 2145, Australia. Phone: (612) 9845 6255. Fax: (612) 9893-8659. E-mail: lyng{at}icpmr.wsahs.nsw.gov.au.

{dagger} Supplemental material for this article may be found at http://aac.asm.org/.

{ddagger} Present address: Department of Microbiology, Royal North Shore Hospital, St. Leonards, NSW, Australia.

Antimicrobial Agents and Chemotherapy, January 2006, p. 204-209, Vol. 50, No. 1
0066-4804/06/$08.00+0     doi:10.1128/AAC.50.1.204-209.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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