Rapid detection of mycobacteria in inflammatory necrotizing granulomas from formalin-fixed, paraffin-embedded tissue by PCR in clinically high-risk patients with acid-fast stain and culture-negative tissue biopsies

William Jackson Hardman, Guy M. Benian, Tom Howard, John E. Mcgowan, Beverly Metchock, James J. Murtagh

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

A collection of inflammatory necrotizing granulomas (INGs) negative by acid-fast stain and culture (AFSC) were analyzed by polymerase chain reaction (PCR) for the presence of mycobacteria. Forty-two paraffin-embedded specimens with INGs were collected from patients at high risk for contracting tuberculosis. Twenty biopsies were positive and 22 were negative for mycobacteria by AFSC. Two universal primers specific for all mycobacteria were used to detected a 414 base pair (bp) fragment of 16S rRNA gene. Twenty of 20 biopsies were positive for mycobacteria by both AFSC and PCR (100%), whereas 19 of 22 biopsies negative by AFSC were positive by PCR (86%). Follow-up of patients who were PCR positive but AFSC negative identified nine patients who had subsequent biopsies. Specimens from eight of these nine patients eventually grew Mycobacterium tuberculosis. Our results demonstrate that the detection of mycobacterial DNA by this method should be used in conjunction with AFSC for the initial diagnosis of mycobacterial infection.

Original languageEnglish (US)
Pages (from-to)384-389
Number of pages6
JournalAmerican Journal of Clinical Pathology
Volume106
Issue number3
DOIs
StatePublished - Jan 1 1996

Fingerprint

Mycobacterium
Granuloma
Paraffin
Formaldehyde
Coloring Agents
Biopsy
Polymerase Chain Reaction
Acids
rRNA Genes
Mycobacterium tuberculosis
Base Pairing
Tuberculosis
DNA
Infection

Keywords

  • 16S rRNA
  • Inflammatory necrotizing granulomas
  • Mycobacterium tuberculosis
  • Polymerase chain reaction
  • Tuberculosis

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Rapid detection of mycobacteria in inflammatory necrotizing granulomas from formalin-fixed, paraffin-embedded tissue by PCR in clinically high-risk patients with acid-fast stain and culture-negative tissue biopsies. / Hardman, William Jackson; Benian, Guy M.; Howard, Tom; Mcgowan, John E.; Metchock, Beverly; Murtagh, James J.

In: American Journal of Clinical Pathology, Vol. 106, No. 3, 01.01.1996, p. 384-389.

Research output: Contribution to journalArticle

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abstract = "A collection of inflammatory necrotizing granulomas (INGs) negative by acid-fast stain and culture (AFSC) were analyzed by polymerase chain reaction (PCR) for the presence of mycobacteria. Forty-two paraffin-embedded specimens with INGs were collected from patients at high risk for contracting tuberculosis. Twenty biopsies were positive and 22 were negative for mycobacteria by AFSC. Two universal primers specific for all mycobacteria were used to detected a 414 base pair (bp) fragment of 16S rRNA gene. Twenty of 20 biopsies were positive for mycobacteria by both AFSC and PCR (100{\%}), whereas 19 of 22 biopsies negative by AFSC were positive by PCR (86{\%}). Follow-up of patients who were PCR positive but AFSC negative identified nine patients who had subsequent biopsies. Specimens from eight of these nine patients eventually grew Mycobacterium tuberculosis. Our results demonstrate that the detection of mycobacterial DNA by this method should be used in conjunction with AFSC for the initial diagnosis of mycobacterial infection.",
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