Use of Trichomonas vaginalis clinical isolates to evaluate correlation of gene expression and metronidazole resistance.
We investigated whether variations in gene expression of enzymes associated with anaerobic resistance of laboratory-derived strains of Trichomonas vaginalis could be detected in a group of 28 clinical isolates with variations in metronidazole sensitivity. We compared isolates by real-time PCR because this method allows for highly sensitive quantification of mRNA and for evaluation of several genes simultaneously. We found that PFOR gene A mRNA levels were highly correlated with PFOR gene B levels, as well as the D subunit of malic enzyme and ferrodoxin. Ferrodoxin mRNA expression was also significantly correlated with that of malic enzyme and hydrogenase. However, when we evaluated relationships between these enzymes and resistance to metronidazole, we found no significant correlations between aerobic or anaerobic in vitro sensitivity to drug and mRNA levels of any of the enzymes tested. Similarly, using a Student's t-test, no significant differences in enzyme mRNA levels were observed between isolates separated by metronidazole resistance or susceptibility. The lack of correlation between gene expression and resistance or susceptibility could be the result of differences in expression at the protein level or because other biochemical pathways or genes are involved in the resistance observed in clinical settings.
Mead
J R
JR
Veterans Affairs Medical Center, Decatur, Georgia 30033, USA. jmead@emory.edu
Fernadez
M
M
Romagnoli
P A
PA
Secor
W E
WE
D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D017931:DNA Primers / Q000737:chemistry; D004351:Drug Resistance / Q000235:genetics*; D005260:Female; D015870:Gene Expression / Q000235:genetics; D020459:Genes, rRNA / Q000235:genetics; D006801:Humans; D006864:Hydrogenase / Q000235:genetics; D008795:Metronidazole / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods; D050743:Pyruvate Synthase / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D013223:Statistics as Topic; D014247:Trichomonas Vaginitis / Q000188:drug therapy*; D014246:Trichomonas vaginalis / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification
D000981:Antiprotozoal Agents; D017931:DNA Primers; D012333:RNA, Messenger; D008795:Metronidazole; D006864:Hydrogenase; D050743:Pyruvate Synthase