Evaluation of PorB variable region typing of Neisseria gonorrhoeae using PCR-ELISA in samples collected from men who have sex with men.
A polymerase chain reaction (PCR)-based enzyme-linked immunosorbent assay (ELISA) methodology was developed to characterize Neisseria gonorrhoeae porB gene variable regions (VR); the methodology was evaluated in comparison to porB VR typing by checkerboard hybridization. Clinical noncultured samples from 35 men who have sex with men (MSM), positive by nucleic amplification assays for N. gonorrhoeae, were typed using a panel of 40 oligonucleotide probes to porB VRs and compared to checkerboard hybridization. Complete concordance was observed between the two methods at PIB VRs 1, 3, and 7. At the more degenerate VRs 5 and 6, PCR ELISA resulted in obtaining more typeable VRs than checkerboard hybridization due to single nucleotide mismatches. By PCR ELISA, two predominant PIB porB types were identified in 58% of the samples and the remaining 16 samples had one of six other porB types. Both PCR ELISA and checkerboard hybridization methods of porB VR typing allowed characterization of N. gonorrhoeae from noncultured clinical samples including throat and rectal swabs and discriminated N. gonorrhoeae from N. meningitidis present in some of the samples. PCR ELISA is a rapid, relatively inexpensive and alternative molecular typing method for N. gonorrhoeae, suitable for use in conjunction with molecular diagnostic tests.
Ling
A E
AE
Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Melbourne, Australia.
Bash
M C
MC
Lynn
F
F
Lister
N A
NA
Zhu
P
P
Garland
S M
SM
Fairley
C K
CK
Tabrizi
S N
SN
D015373:Bacterial Typing Techniques; D001483:Base Sequence; D015342:DNA Probes / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D014644:Genetic Variation; D006069:Gonorrhea / Q000175:diagnosis / Q000382:microbiology*; D018451:Homosexuality, Male; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D018272:Porins / Q000145:classification / Q000235:genetics* / Q000276:immunology
D015342:DNA Probes; D004269:DNA, Bacterial; D018272:Porins; C057699:porin protein, Neisseria