One-step reverse transcriptase PCR method for detection of Borrelia burgdorferi mRNA in mouse Lyme arthritis tissue samples.
A one-step reverse transcriptase PCR (RT-PCR) method for detection of Borrelia burgdorferi mRNA in infected C3H mice is described. This simple procedure, less prone to nucleic acid cross-contamination than the standard method, was found to be 10-fold more sensitive than a classical two-step RT-PCR assay. By using one-step RT-PCR, flagellin mRNAs were detected in synovial and heart tissues from all seven infected mice tested.
D000818:Animals; D001170:Arthritis, Infectious / Q000209:etiology* / Q000821:virology; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000302:isolation & purification*; D005408:Flagellin / Q000235:genetics; D006321:Heart / Q000821:virology; D008193:Lyme Disease / Q000150:complications* / Q000175:diagnosis*; D051379:Mice; D008809:Mice, Inbred C3H; D010493:Pericarditis / Q000209:etiology / Q000821:virology; D012333:RNA, Messenger / Q000032:analysis*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013583:Synovial Membrane / Q000821:virology