title: A multiplex touchdown PCR for detection of Streptococcus pneumoniae, Haemophilus influenzae type b and Mycobacterium tuberculosis complex in sputum samples.
abstract: Rapid and accurate detection of Streptococcus pneumoniae (Sp), Haemophilus   influenzae type b (Hib) and Mycobacterium tuberculosis complex (MTBC) in sputum by   conventional methods remains problematic. Primers based on capsular polysaccharide   biosynthesis gene (cpsA), the region II of the capsulation locus (cap), the insertion sequence   IS6110 were designed for Sp, Hib, MTBC respectively. These primers were incorporated in a   multiplex touchdown PCR assay for simultaneous detection of Sp, Hib and MTBC. The   multiplex touchdown PCR assay was evaluated using standard strains and clinical sputum   samples. The multiplex touchdown PCR assay showed 100% specificity in identifying Sp, Hib,   MTBC from pure culture of standard strains. The sensitivities of the multiplex touchdown   PCR assay were 94%, 98%, 98% for detection of Sp, Hib and MTBC respectively based on   culture results while evaluated using 492 consecutive qualified clinical sputum samples; the   specificities were all 100%. This highly sensitive and specific multiplex touchdown PCR   assay offers a rapid and simple method for detection of Sp, Hib and MTBC in clinical sputum   samples.
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212013, China.
authors.forename: Y C
authors.initials: YC
authors.lastname: Luo
authors.forename: P
authors.initials: P
authors.lastname: Du
authors.forename: J Z
authors.initials: JZ
authors.lastname: Zhao
authors.forename: X J
authors.initials: XJ
authors.lastname: Duan
authors.forename: Y J
authors.initials: YJ
authors.lastname: Hou
authors.forename: H
authors.initials: H
authors.lastname: Pan
authors.forename: S H
authors.initials: SH
authors.lastname: Shao
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D006192:Haemophilus Infections / Q000175:diagnosis / Q000382:microbiology; D019741:Haemophilus influenzae type b / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification*; D011008:Pneumococcal Infections / Q000175:diagnosis / Q000382:microbiology; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013183:Sputum / Q000382:microbiology*; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D014376:Tuberculosis / Q000175:diagnosis / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; C500439:CpsA protein, Streptococcus pneumoniae; D017931:DNA Primers; D004269:DNA, Bacterial