title: Performance characteristics of nested polymerase chain reaction vs real-time polymerase chain reaction methods for detecting Mycobacterium tuberculosis complex in paraffin-embedded human tissues.
abstract: OBJECTIVE
  Nucleic acid amplification tests on formalin-fixed, paraffin-embedded (FFPE) tissue specimens enable Mycobacterium tuberculosis complex (MTB) detection and rapid tuberculosis diagnosis in the absence of microbiologic culture tests. We aimed to evaluate the efficacy of different polymerase chain reaction (PCR) methods for detecting Mycobacterium species in FFPE tissues.
  
  
  METHODS
  We examined 110 FFPE specimens (56 nonmycobacterial cases, 32 MTB, and 22 nontuberculous mycobacteria [NTM] determined by acid-fast bacilli [AFB] culture) to assess five PCR methods: nested PCR (N-PCR) (Seeplex MTB Nested ACE Detection; Seegene, Seoul, South Korea), an in-house real-time PCR (RT-PCR) method, and three commercial RT-PCR methods (AccuPower MTB RT-PCR [Bioneer, Seoul, Korea], artus M tuberculosis TM PCR [Qiagen, Hilden, Germany], and AdvanSure tuberculosis/NTM RT-PCR [LG Life Sciences, Seoul, Korea]).
  
  
  RESULTS
  The results of N-PCR, in-house RT-PCR, and AdvanSure RT-PCR correlated well with AFB culture results (concordance rates, 94.3%, 87.5%, and 89.5%, respectively). The sensitivity of N-PCR (87.5%) was higher than that of the RT-PCR methods, although these differences were not statistically significant between N-PCR and the in-house and AdvanSure RT-PCR methods (68.8% and 80.0%, respectively). All the PCR methods had high specificities, ranging from 98.2% to 100%. Only two NTM cases were detected by AdvanSure RT-PCR, implying a very low sensitivity.
  
  
  CONCLUSIONS
  Well-designed RT-PCR and N-PCR can effectively identify MTB in FFPE specimens.
authors.affiliation: From the Department of Pathology, Seoul National University Bundang Hospital, Seongnam, Gyeonggido, Republic of Korea; Department of Pathology, Kyungpook National University School of Medicine, Jung-gu, Daegu, Republic of Korea;
authors.forename: An Na
authors.initials: AN
authors.lastname: Seo
authors.affiliation: From the Department of Pathology, Seoul National University Bundang Hospital, Seongnam, Gyeonggido, Republic of Korea;
authors.forename: Hyo Jin
authors.initials: HJ
authors.lastname: Park
authors.affiliation: From the Department of Pathology, Seoul National University Bundang Hospital, Seongnam, Gyeonggido, Republic of Korea;
authors.forename: Hye Seung
authors.initials: HS
authors.lastname: Lee
authors.affiliation: From the Department of Pathology, Seoul National University Bundang Hospital, Seongnam, Gyeonggido, Republic of Korea;
authors.forename: Jung Ok
authors.initials: JO
authors.lastname: Park
authors.affiliation: Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Gyeonggido, Republic of Korea; and.
authors.forename: Ho Eun
authors.initials: HE
authors.lastname: Chang
authors.affiliation: From the Department of Pathology, Seoul National University Bundang Hospital, Seongnam, Gyeonggido, Republic of Korea;
authors.forename: Kyung Han
authors.initials: KH
authors.lastname: Nam
authors.affiliation: From the Department of Pathology, Seoul National University Bundang Hospital, Seongnam, Gyeonggido, Republic of Korea; Departments of Pathology and.
authors.forename: Gheeyoung
authors.initials: G
authors.lastname: Choe
authors.affiliation: Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Gyeonggido, Republic of Korea; and Laboratory Medicine, Seoul National University College of Medicine, Jongnogu, Seoul, Republic of Korea. m91w95pf@snu.ac.kr.
authors.forename: Kyoung Un
authors.initials: KU
authors.lastname: Park
mesh_terms: D006801:Humans; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity
keywords: Formalin-fixed; Mycobacterium tuberculosis; Nested PCR; Real-time PCR; paraffin-embedded tissues