rpoB nested PCR and sequencing for the early diagnosis of tuberculous meningitis and rifampicin resistance.
METHODS
The prognosis of tuberculous meningitis (TBM) is linked to early diagnosis and prescription of adequate treatment.


OBJECTIVE
To evaluate the efficacy of the rpoB nested polymerase chain reaction (PCR) and sequencing assay to detect and identify Mycobacterium tuberculosis complex (MTC) strains and strains resistant to rifampicin (RMP) in cerebrospinal fluid specimens (CSF) from patients with highly suspected TBM.


METHODS
Retrospective blinded hospital-based study.


RESULTS
rpoB nested PCR and sequencing assay detected MTC in 31/36 CSF specimens from 16 patients with clinically suspected TBM. All of the control CSF specimens from 25 patients with non-TBM showed negative results. One of the 16 patients had a mutation at codon C526G as compared to the rpoB sequences in GenBank. This corresponds to a diagnostic sensitivity of 86% (95%CI 71-95) and a specificity of 100% (95%CI 86-100).


CONCLUSIONS
Our results suggest that rpoB nested PCR and sequencing assay can detect MTC and simultaneously determine its RMP susceptibility in CSF from patients with highly suspected TBM.
Huang
H-J
HJ
State Key Laboratory for Diagnosis and Treatment of Infectious Disease, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
Xiang
D-R
DR
Sheng
J-F
JF
Li
J
J
Pan
X-P
XP
Yu
H-Y
HY
Sheng
G-P
GP
Li
Lan-Juan
LJ
D000293:Adolescent; D000328:Adult; D000368:Aged; D000995:Antitubercular Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000134:cerebrospinal fluid / Q000235:genetics; D002555:Cerebrospinal Fluid / Q000382:microbiology; D004269:DNA, Bacterial / Q000134:cerebrospinal fluid / Q000235:genetics; D012321:DNA-Directed RNA Polymerases; D024881:Drug Resistance, Bacterial / Q000187:drug effects / Q000235:genetics*; D042241:Early Diagnosis; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D012293:Rifampin / Q000494:pharmacology; D012680:Sensitivity and Specificity; D016037:Single-Blind Method; D014390:Tuberculosis, Meningeal / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics* / Q000382:microbiology; D055815:Young Adult
D000995:Antitubercular Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C505805:rpoB protein, Mycobacterium tuberculosis; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin