title: Sequencing-based detection of drug-resistant Mycobacterium tuberculosis in patients with spinal tuberculosis.
abstract: BACKGROUND
  Traditionally, bacterial cultures serve as the gold standard for the detection of drug resistance and can provide evidence for directing the treatment of tuberculosis. However, this method has a low positive rate and is time consuming, which significantly limits its wide application. Thus, in the present study, the genes associated with drug resistance were amplified and sequenced to determine the presence of drug-resistant mutations in Mycobacterium tuberculosis. In addition, to find a more sensitive, specific, rapid, and simple method for the detection of drug-resistant bacteria, this method was compared with traditional bacterial culture.
  
  
  METHODS
  Pus was collected from surgical patients with spinal tuberculosis. The common drug resistance genes (rpoB, rpsL, and katG) were amplified by PCR. The PCR products were then sequenced, and the sequences were compared with those in the NCBI database using DNATools v.5.1 software.
  
  
  RESULTS
  Mutations were identified in 17 patients, including mutations in the rpsL gene in four patients, the rpoB gene in seven, and the katG gene in six. The mean time of detection was 6 days.
  
  
  CONCLUSIONS
  These results indicated that PCR and DNA sequencing are rapid, sensitive, and specific methods for the detection of drug-resistant genes of M. tuberculosis in patients with spinal tuberculosis. This method may provide critical evidence for the clinical treatment of tuberculosis when it is applied in combination with bacterial culture.
authors.affiliation: General Hospital of Ningxia Medical University, No. 804 Shengli St., Yinchuan, Ningxia, China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Si
authors.forename: Zhe
authors.initials: Z
authors.lastname: Wang
authors.forename: Zili
authors.initials: Z
authors.lastname: Wang
authors.forename: Haomin
authors.initials: H
authors.lastname: Li
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000235:genetics*; D002374:Catalase / Q000235:genetics; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial; D012321:DNA-Directed RNA Polymerases; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000502:physiology; D016133:Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D013997:Time Factors; D014399:Tuberculosis, Spinal / Q000382:microbiology*; D055815:Young Adult
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012269:Ribosomal Proteins; C505805:rpoB protein, Mycobacterium tuberculosis; D002374:Catalase; C480353:katG protein, Mycobacterium tuberculosis; D012321:DNA-Directed RNA Polymerases