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Real-Time PCR AssayUsing Fine-NeedleAspirates and TissueBiopsy Specimens forRapid Diagnosis ofMycobacterialLymphadenitis inChildren E. S. Bruijnesteijn vanCoppenraetJerôme A. LindeboomJan M. Prins2004 год

Real-Time PCR Assay Using Fine-Needle Aspirates and Tissue Biopsy Specimens for Rapid Diagnosis of Mycobacterial Lymphadenitis in Children
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Аннотация: ABSTRACT A real-time PCR assay was developed to diagnose and identify the causative agents of suspected mycobacterial lymphadenitis. Primers and probes for the real-time PCR were designed on the basis of the internal transcribed spacer sequence, enabling the recognition of the genus Mycobacterium and the species Mycobacterium avium and M . tuberculosis . The detection limit for the assay was established at 1,100 CFU/ml of pus, and the specificity tests showed no false-positive reaction with other mycobacterial species and other pathogens causing lymphadenitis. From 67 children with suspected mycobacterial lymphadenitis based on a positive mycobacterial skin test, 102 samples (58 fine-needle aspirates [FNA] and 44 tissue specimens) were obtained. The real-time PCR assay detected a mycobacterial infection in 48 patients (71.6%), whereas auramine staining and culturing were positive for 31 (46.3%) and 28 (41.8%) of the patients. The addition of the real-time PCR assay to conventional diagnostic tests resulted in the recognition of 13 more patients with mycobacterial disease. These results indicate that the real-time PCR is more sensitive than conventional staining and culturing techniques ( P = 0.006). The M . avium -specific real-time PCR was positive for 38 patients, and the M . tuberculosis -specific real-time PCR was positive for 1 patient. Analysis of 27 patients from whom FNA and tissue biopsy specimens were collected revealed significantly more positive real-time PCR results for FNA than for tissue biopsy specimens ( P = 0.003). Samples from an age-matched control group of 50 patients with PCR-proven cat scratch disease were all found to be negative by the real-time PCR. We conclude that this real-time PCR assay with a sensitivity of 72% for patients with lymphadenitis and a specificity of 100% for the detection of atypical mycobacteria can provide excellent support for clinical decision making in children with lymphadenitis.
Год издания: 2004
Авторы: E. S. Bruijnesteijn van Coppenraet, Jerôme A. Lindeboom, Jan M. Prins, Marc Peeters, Eric C. J. Claas, Ed J. Kuijper
Издательство: American Society for Microbiology
Источник: Journal of Clinical Microbiology
Ключевые слова: Mycobacterium research and diagnosis, Bartonella species infections research, Infectious Diseases and Tuberculosis
Другие ссылки: Journal of Clinical Microbiology (HTML)
Europe PMC (PubMed Central) (PDF)
Europe PMC (PubMed Central) (HTML)
PubMed Central (HTML)
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