PURPOSE: PCR is widely used for rapidly and accurately detecting Mycobacterium Species. The purpose of this study was to assess the diagnostic performance of three real-time PCR kits and evaluate the concordance with two older PCR methods. MATERIALS AND METHODS: Using 128 samples, the five PCR methods were assessed, including an in-house PCR protocol, the COBAS Amplicor MTB, the COBAS TaqMan MTB, the AdvanSure TB/NTM real-time PCR, and the Real-Q M. tuberculosis kit. The discrepant results were further examined by DNA sequencing and using the AdvanSure Mycobacteria Genotyping Chip for complete analysis. RESULTS: For Mycobacterium tuberculosis (MTB) detection, all five kits showed 100% matching results (positive; N = 11 and negative; N = 80). In non-tuberculous mycobacterium (NTM) discrimination, the AdvanSure yielded two true-positive outcomes from M. intracellulare and one false positive outcome, while the Real-Q resulted in one true-positive outcome and one false negative outcome for each case and another false negative result using the provided DNA samples. CONCLUSION: Real-time PCR, yielded results that were comparable to those of the older PCR methods for detecting MTB. However, there were disagreements among the applied kits in regard to the sample test results for detecting NTM. Therefore, we recommend that additional confirmatory measures such as DNA sequencing should be implemented in such cases, and further research with using a larger numbers of samples is warranted to improve the detection of NTM.
DNA, Bacterial/genetics ; Humans ; Mycobacterium/*genetics ; Mycobacterium Infections/*diagnosis/microbiology ; Mycobacterium avium Complex/genetics ; Mycobacterium avium-intracellulare Infection/diagnosis ; Mycobacterium tuberculosis/genetics ; Polymerase Chain Reaction/*standards ; Reagent Kits, Diagnostic/*standards ; Tuberculosis/diagnosis

INTRODUCTIONGranulomatous cervicofacial lymphadenitis (GCL) is not uncommon in children. Nontuberculous mycobacteria (NTM) seem to be the predominant cause. We sought to study the clinical and microbiological profile of patients with GCL, and identify features that may impact outcome.

METHODSChildren aged < 16 years who presented to KK Women's and Children's Hospital, Singapore, between January 1998 and December 2006, and who had GCL were identified from laboratory records. Clinical and laboratory data was collected and analysed for risk factors for patients with positive lymph node cultures, and for patients with and without recurrence after treatment.

RESULTSIn all, 60 children were identified, with a median age of 56 (interquartile range [IQR] 34-101) months. Median duration of symptoms before presentation was 5 (IQR 4-8) weeks. The majority presented with single (73.3%) or unilateral (96.7%) lymphadenopathy, located in the submandibular, preauricular/parotid or infra-/post-auricular region (76.7%). Out of 51 patients, 26 (51.0%) had a tuberculin skin test reading of ≥ 10 mm. Out of 52 patients, 10 (19.2%) had positive mycobacterial cultures, which included seven isolates of NTM. Out of 34 cases, tuberculous polymerase chain reaction was positive in 11 (32.4%). With regard to recurrence after initial treatment, age < 5 years at presentation was found to be a predictor for recurrence (p = 0.008), while initial complete excision of affected nodes predicted no recurrence (p = 0.003).

CONCLUSIONIn our study, younger age was noted to be associated with a higher chance of recurrence, while complete excision of the involved node at initial presentation predicted non-recurrence.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Lymphadenitis ; diagnosis ; epidemiology ; microbiology ; Lymphatic Diseases ; diagnosis ; microbiology ; Male ; Mycobacterium ; Mycobacterium Infections ; diagnosis ; epidemiology ; microbiology ; Recurrence ; Retrospective Studies ; Singapore ; Treatment Outcome ; Tuberculin Test

The incidence of pulmonary disease caused by nontuberculous mycobacteria (NTM) appears to be increasing worldwide. In Korea, M. avium complex and M. abscessus account for most of the pathogens encountered, whilst M. kansasii is a relatively uncommon cause of NTM pulmonary diseases. NTM pulmonary disease is highly complex in terms of its clinical presentation and management. Because its clinical features are indistinguishable from those of pulmonary tuberculosis and NTMs are ubiquitous in the environment, the isolation and identification of causative organisms are mandatory for diagnosis, and some specific diagnostic criteria have been proposed. The treatment of NTM pulmonary disease depends on the infecting species, but decisions concerning the institution of treatment are never easy. Treatment requires the use of multiple drugs for 18 to 24 months. Thus, treatment is expensive, often has significant side effects, and is frequently not curative. Therefore, clinicians should be confident that there is sufficient pathology to warrant prolonged, multidrug treatment regimens. In all of the situations, outcomes can be best optimized only when clinicians, radiologists, and laboratories work cooperatively.
Anti-Bacterial Agents/therapeutic use ; Female ; Humans ; Korea ; Lung Diseases/*diagnosis/drug therapy/microbiology ; Male ; Mycobacterium Infections, Atypical/*diagnosis/drug therapy/microbiology ; Mycobacterium avium-intracellulare Infection/diagnosis/drug therapy/microbiology ; Research Support, Non-U.S. Gov't

BACKGROUND: The combined use of liquid media and solid media is recommended for mycobacterial culture. We evaluated diagnostic performance of combination of BACTEC Mycobacteria Growth Indicator Tube (MGIT; Becton Dickinson, USA) and 2% Ogawa media (Korean Institute of Tuberculosis, Korea) for recovery of mycobacteria. METHODS: In September 2007, 1,764 specimens from 1,059 patients were cultured with MGIT and Ogawa. Acid fast bacilli (AFB) smear was fluorochrome-stained. The isolates were identified into Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM) with PCR using Seeplex TB Detection Kit (Seegene, Korea). Recovery rate, time to detection (TTD), contamination rate, mixed growth rate and species distribution were analyzed. RESULTS: Two hundred thirty-five specimens (13.3%) from 165 patients (15.6%) were positive for mycobacterial culture. Recovery rates of mycobacteria from the group using both media, MGIT only, and Ogawa only were 13.3%, 12.1%, and 7.8%, respectively. While MGIT recovered 98.9% of MTB and 79.7% of NTM, Ogawa recovered 65.9% of MTB and 54.1% of NTM. TTDs of total mycobacteria/MTB/NTM in MGIT and Ogawa were 10.6/11.4/9.7 days and 31/29/33 days, respectively. MGIT TTDs of total mycobacteria/MTB/NTM from AFB-positive specimens were significantly shorter than those of AFB-negative specimens; 8.2/9.5/4.4 days vs 11.6/12.7/10.7 days. Contamination and mixed growth rate of MGIT were 9.6% and 3.7%. Primary culture of Ogawa recovered 1 MTB and 1 NTM among the 170 MGIT-contaminated specimens and 38 mycobacteria among 66 specimens that showed mixed cultures of MGIT. CONCLUSIONS: MGIT warrants sensitive and rapid isolation of mycobacteria. However, the combination of MGIT and Ogawa is more desirable to recover mycobacteria in the case of contaminations or mixed cultures.
*Culture Media ; False Positive Reactions ; Humans ; Mycobacterium/*growth & development/isolation & purification ; Mycobacterium Infections/*diagnosis/microbiology ; Mycobacterium tuberculosis/*growth & development/isolation & purification ; Reagent Kits, Diagnostic ; Sensitivity and Specificity ; Sputum/microbiology ; Time Factors

We used HPLC and AdvanSure real-time PCR (LG Life Sciences, Korea) to retrospectively analyze non-tuberculous mycobacteria (NTM) in 133 clinical specimens. The specimens were culture-positive for NTM and the HPLC method identified 130 strains of mycobacteria from the cultures (97.7%) at the species level. Among the isolates, 48 Mycobacterium. kansasii (36.1%), 39 M. intracellulare (29.3%), 17 M. avium (12.8%), 16 M. abscessus (12.0%), 6 M. fortuitum (4.5%), 2 M. szulgai (1.5%), 2 M. gordonae (1.5%), and 3 unclassified NTM strains (2.3%) were identified. The real-time PCR assay identified 60 NTM-positive specimens (45.1%), 65 negative specimens (48.9%), and 8 M. tuberculosis (TB)-positive specimens (6.0%). The real-time PCR assay is advantageous because of its rapid identification of NTM. However, in our study, the real-time PCR assay showed relatively low sensitivity (45.1%) when using direct specimens including sputum and bronchoalveolar lavage (BAL) fluid. HPLC is useful as it discriminates NTM at the species level, although it is time-consuming and requires specific equipment and technical expertise. A combination of both methods will be helpful for the rapid and accurate identification of mycobacteria in clinical laboratories.
Aged ; Aged, 80 and over ; Bronchoalveolar Lavage Fluid/microbiology ; *Chromatography, High Pressure Liquid ; DNA, Bacterial/genetics ; Female ; Humans ; Male ; Middle Aged ; Mycobacterium/*genetics/isolation & purification ; Mycobacterium Infections/diagnosis/*microbiology ; *Real-Time Polymerase Chain Reaction ; Sputum/microbiology

We herein report a case in which the recently characterized species Mycobacterium monacense was isolated from the sputum of an Iranian patient. This case represents the first isolation of M. monacense from Iran. The isolate was identified by conventional and molecular techniques. Our findings show that M. monacense infection is not restricted to developed countries.
Bacterial Proteins/genetics ; Chaperonin 60/genetics ; Chronic Disease ; Female ; Humans ; Iran ; Lung Diseases/diagnosis/*microbiology ; Middle Aged ; Mycobacterium/classification/*genetics/isolation & purification ; Mycobacterium Infections/*microbiology/pathology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Sputum/microbiology

Nontuberculous mycobacterial infections can cause destructive tenosynovitis of the hand. We report on and discuss the clinical course and distinctive radiologic findings of two patients with hand tenosynovitis secondary to M. marinum and intracellulare infection, which are different from those of the nontuberculous mycobacterial infections reported in the previous literature.
Female ; *Hand/radiography ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Mycobacterium Infections, Nontuberculous/*diagnosis/etiology/radiography ; Mycobacterium avium-intracellulare Infection/*diagnosis/etiology/radiography ; *Mycobacterium marinum ; Surgical Wound Infection/complications ; Tenosynovitis/diagnosis/*microbiology/radiography ; Wound Infection/complications

Mycobacterium xenopi is a nontuberculous mycobacterium (NTM) that rarely causes pulmonary disease in Asia. Here we describe the first case of M. xenopi pulmonary disease in Korea. A 66-year-old man was admitted to our hospital with a 2-month history of productive cough and hemoptysis. His past medical history included pulmonary tuberculosis 44 years earlier, leading to a right upper lobectomy. Chest X-ray upon admission revealed cavitary consolidation involving the entire right lung. Numerous acid-fast bacilli were seen in his initial sputum, and M. xenopi was subsequently identified in more than five sputum cultures, using molecular methods. Despite treatment with clarithromycin, rifampicin, ethambutol, and streptomycin, the infiltrative shadow revealed on chest X-ray increased in size. The patient's condition worsened, and a right completion pneumonectomy was performed. The patient consequently died of respiratory failure on postoperative day 47, secondary to the development of a late bronchopleural fistula. This case serves as a reminder to clinicians that the incidence of NTM infection is increasing in Korea and that unusual NTM are capable of causing disease in non-immunocompromised patients.
Aged ; Bacterial Proteins/genetics ; Heat-Shock Proteins/genetics ; Humans ; Korea ; Lung Diseases/*diagnosis/*microbiology/radiography ; Male ; Mycobacterium Infections, Atypical/*diagnosis/microbiology/radiography ; Mycobacterium xenopi/classification/genetics/*isolation & purification ; Phylogeny ; Sequence Analysis, DNA

No abstract available.
Aged ; Bronchiectasis/*diagnosis/microbiology ; Humans ; Male ; Mycobacterium/classification/*genetics/isolation & purification ; Mycobacterium Infections/*diagnosis/microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/chemistry/genetics/metabolism ; Republic of Korea ; Sequence Analysis, DNA

Mycobacterium neoaurum is rapidly growing mycobacteria that can cause human infections. It commonly causes bloodstream infections in immunocompromised hosts, and unlike other mycobacteria species, it rarely causes pulmonary infections. We confirmed the first pulmonary infection case in Korea caused by M. neoaurum using full-length 16S rRNA gene sequencing.
Adult ; Female ; Humans ; Lung Diseases/*diagnosis/microbiology ; Mycobacterium/genetics/*isolation & purification ; Mycobacterium Infections/*diagnosis/microbiology ; Nontuberculous Mycobacteria/genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Republic of Korea ; Sequence Analysis, RNA