Am. J. Respir. Crit. Care Med.,
Volume 161, Number 3, March 2000, 930-934
Sensitive Identification of Mycobacterial Species
Using PCR-RFLP on Bronchial Washings
EIKO
HIDAKA,
TAKAYUKI
HONDA,
ICHIRO
UENO,
YOSHITAKA
YAMASAKI,
KEISHI
KUBO,
and
TSUTOMU
KATSUYAMA
Departments of Laboratory Medicine and Internal Medicine, Shinshu University School of Medicine, Matsumoto, Japan
In 98 patients (24 with active pulmonary tuberculosis [TB] lesions, 28 with cured TB lesions, and 46 with nontuberculous opacities [control group] in chest CT scans), we examined whether washing the
bronchus after brushing the lesion, then applying polymerase chain reaction-restriction fragment
length polymorphism (PCR-RFLP) to the bronchial washings might be useful for diagnosing TB and nontuberculous mycobacteriosis (NTMosis). After biopsy and brushing with a bronchoscope, the bronchus connecting to the lesion was washed with 20 ml saline. The saline used for washing the brushes (5 ml; brushing sample), and 3 to 10 ml saline aspirated through the forceps channel (washing sample) were examined by PCR-RFLP, which proved able to identify Mycobacterium tuberculosis and seven species of nontuberculous mycobacteria (NTM). The values obtained for the sensitivity of the PCR-RFLP with respect to the brushing sample, the washing sample, and both samples mixed together
were 70, 76, and 91%, respectively, when only patients who were culture-positive or radiologically
improved after antituberculous therapy were considered as showing true infection. A mixture of
brushing and washing samples provides useful material for PCR and culture, and the PCR-RFLP used
here is a good method for the simultaneous identification of several species of mycobacterium (including M. tuberculosis). Hidaka E, Honda T, Ueno I, Yamasaki Y, Kubo K, Katsuyama T. Sensitive identification of mycobacterial species using PCR-RFLP on bronchial washings.
