title: [Validation of the PCR-based universal heteroduplex generator assay for identification of rifampicin-resistant and multiresistant Mycobacterium tuberculosis in Lima, Peru].
abstract: OBJECTIVE
  To validate the polymerase chain reaction-based universal heteroduplex generator (PCR UHG-Rif) assay for identifying rifampin-resistant and multidrug-resistant (MDR, resistant to isoniazid and rifampin) Mycobacterium tuberculosis in patients with pulmonary tuberculosis from communities in Lima (Peru) with a high incidence of resistant tuberculosis.
  
  
  METHODS
  To compare the results of antituberculosis drug susceptibility testing in clinical samples performed by the proportion method with those obtained by the PCR UHG-Rif assay, with the aim of analyzing the diagnostic capability of PCR UHG-Rif.
  
  
  RESULTS
  Concordance for the identification of antituberculosis drug susceptibility was 0.95 (kappa = 0.899; P < .05), with a sensitivity and specificity of 0.973 and 0.922 (P < .05), respectively. The positive predictive value was 0.939 (95% CI: 0.879-0.970) and the negative predictive value was 0.965 (95% CI: 0.902-0.988). Nevertheless, the probability for MDR prediction was 0.981 (P < .05). PCR UHG-Rif allows the detection of mixed populations; discordant results can be explained by the presence of point mutations, missense mutations and mutations outside the rpoB "hot" region associated with rifampin-resistance.
  
  
  CONCLUSIONS
  The PCR UHG-Rif assay detects mutations in the rpoB gene with excellent sensitivity and specificity, and suitable predictive values when compared with the standard method for determining susceptibility to antituberculosis drugs. This test can be considered an excellent tool that can contribute to tuberculosis control by correctly identifying patients infected with resistant and MDR bacilli, leading to a reduction in the cases of tuberculosis and resistant tuberculosis.
authors.affiliation: Laboratorio de Biotecnología y Biología Molecular, Instituto Nacional de Salud, Lima, Peru. rcalderon@ins.gob.pe
authors.forename: Róger I
authors.initials: RI
authors.lastname: Calderón-Espinoza
authors.forename: Luis
authors.initials: L
authors.lastname: Asencios-Solís
authors.forename: Neyda
authors.initials: N
authors.lastname: Quispe-Torres
authors.forename: Gloria
authors.initials: G
authors.lastname: Yale-Cajahuanca
authors.forename: Carmen
authors.initials: C
authors.lastname: Suárez-Nole
authors.forename: Leonid
authors.initials: L
authors.lastname: Lecca-García
authors.forename: Luis F
authors.initials: LF
authors.lastname: Llanos-Zavalaga
mesh_terms: D000904:Antibiotics, Antitubercular / Q000627:therapeutic use*; D004311:Double-Blind Method; D004352:Drug Resistance, Microbial; D020180:Heteroduplex Analysis; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification*; D010568:Peru; D016133:Polymerase Chain Reaction; D011446:Prospective Studies; D012293:Rifampin / Q000627:therapeutic use*; D018088:Tuberculosis, Multidrug-Resistant / Q000382:microbiology*
chemical_list: D000904:Antibiotics, Antitubercular; D012293:Rifampin