Evaluation of acridinium-ester-labeled DNA probes for identification of Mycobacterium tuberculosis and Mycobacterium avium-Mycobacterium intracellulare complex in culture

Goto, Mieko; Oka, Shinichi; Okuzumi, Katsuko; Kimura, Satoshi; Shimada, Kaoru

doi:10.1128/jcm.29.11.2473-2476.1991

Cited by 118 publications

(38 citation statements)

References 15 publications

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“…Identification of mycobacteria. Routine biochemical methods (19), gas-liquid chromatography (25), and the Accuprobe culture confirmation tests (Gen-Probe Inc., San Diego, Calif.) (18) were used for isolate identification.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of the semiautomated Abbott LCx Mycobacterium tuberculosis assay for direct detection of Mycobacterium tuberculosis in respiratory specimens

et al. 1997

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Five hundred twenty processed respiratory specimens from 326 patients received for the diagnosis of tuberculosis or other mycobacterial infections were tested by means of the LCx Mycobacterium tuberculosis Assay from Abbott Laboratories, which uses ligase chain reaction technology for the direct detection of M. tuberculosis complex in respiratory specimens. The results of the LCx M. tuberculosis Assay were compared with the results of culture and staining techniques. After a combination of culture results and the patient's clinical data, a total of 195 specimens were collected from 110 patients who were positively diagnosed as having pulmonary tuberculosis. Twenty-three of these 195 specimens which corresponded to 10 patients with a history of pulmonary tuberculosis (TB) and anti-TB treatment ranging from 1 to 6 months were culture negative. The other 172 specimens were culture positive for M. tuberculosis. With an overall positivity rate of 37.5% (195 of 520 specimens), the sensitivity, specificity, and positive and negative predictive values were 90.8, 100, 100, and 94.7%, respectively, for the LCx M. tuberculosis Assay; 88.2, 100, 100, and 93.4%, respectively, for culture; and 82.6, 92, 72.9, and 97.6%, respectively, for acid-fast staining. For 161 specimens (82.6%) from patients smear positive for the disease and 34 specimens (17.4%) from patients smear negative for the disease, the sensitivity values for the LCx M. tuberculosis Assay were 98.8 and 53%, respectively. There were no statistically significant differences in the sensitivities and specificities between the LCx M. tuberculosis Assay and culture (P > 0.05). Conclusively, the LCx M. tuberculosis Assay has proved to have an acceptable sensitivity and a high specificity in detecting M. tuberculosis and has the potential of reducing the diagnosis time to an 8-h working day.Semiquantitative reporting modified from a previous report (22); 0, no acid-fast bacilli seen; 1ϩ, a few acid-fast bacilli seen; 2ϩ, moderate numbers of acid-fast bacilli seen; 3ϩ, many acid-fast bacilli seen. c

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Section: Methodsmentioning

confidence: 99%

Evaluation of the semiautomated Abbott LCx Mycobacterium tuberculosis assay for direct detection of Mycobacterium tuberculosis in respiratory specimens

et al. 1997

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“…By conventional biochemical methods (52,57), the identification of the species to which the cultured mycobacterial strain belongs may require an additional 2 to 4 weeks. More rapid methods for the identification of cultured mycobacteria are the analysis of lipid composition (7,11,33), the use of species-specific antibodies (44,56), and species-specific DNA or RNA probes (14,16,32).…”

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confidence: 99%

PCR assay based on DNA coding for 16S rRNA for detection and identification of mycobacteria in clinical samples

et al. 1995

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A PCR and a reverse cross blot hybridization assay were developed for the detection and identification of mycobacteria in clinical samples. The PCR amplifies a part of the DNA coding for 16S rRNA with a set of primers that is specific for the genus Mycobacterium and that flanks species-specific sequences within the genes coding for 16S rRNA. The PCR product is analyzed in a reverse cross blot hybridization assay with probes specific for M. tuberculosis complex (pTub1), M. avium (pAvi3), M. intracellulare (pInt5 and pInt7), M. kansasii complex-M. scrofulaceum complex (pKan1), M. xenopi (pXen1), M. fortuitum (pFor1), M. smegmatis (pSme1), and Mycobacterium spp. (pMyc5a). The PCR assay can detect 10 fg of DNA, the equivalent of two mycobacteria. The specificities of the probes were tested with 108 mycobacterial strains (33 species) and 31 nonmycobacterial strains (of 17 genera). The probes pAvi3, pInt5, pInt7, pKan1, pXen1, and pMyc5a were specific. With probes pTub1, pFor1, and pSme1, slight cross hybridization occurred. However, the mycobacterial strains from which the cross-hybridizing PCR products were derived belonged to nonpathogenic or nonopportunistic species which do not occur in clinical samples. The test was used on 31 different clinical specimens obtained from patients suspected of having mycobacterial disease, including a patient with a double mycobacterial infection. The samples included sputum, bronchoalveolar lavage, tissue biopsy samples, cerebrospinal fluid, pus, peritoneal fluid, pleural fluid, and blood. The results of the PCR assay agreed with those of conventional identification methods or with clinical data, showing that the test can be used for the direct and rapid detection and identification of mycobacteria in clinical samples.

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“…9 Demonstration of acid-fast bacilli (AFB) on microscopy and their recovery in culture on Lowenstein±Jensen (LJ) medium or by guinea pig inoculation are also disappointing in most instances, probably due to the presence of highly attenuated or nonviable bacilli in this form of cutaneous tuberculosis or the fact that the skin lesions represent a reaction pattern to the antigenic component of the bacilli in the sensitized host. 10,11 Therefore, more speci®c and rapid diagnostic modalities, such as polymerase chain reaction (PCR), 12 radioimmunoassay (RIA), and 125 I-or acridineester-labeled DNA probes, 13 are being developed, but at present the clinician has to rely on clinical and histopathologic correlation for the diagnosis.…”

Section: Discussionmentioning

confidence: 99%

An unusual episode of lupus vulgaris masquerading as sporotrichosis

2001

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Case 1 A 24‐year‐old man presented with asymptomatic, gradually progressive plaques and nodules over the right knee extending to the thigh in a linear pattern of 3 months' duration. A year previously, he had been diagnosed with tuberculosis of the spine and had been advised to take antitubercular treatment; however, he chose to stop treatment on his own after 4 months of therapy as his spinal condition improved with a marked decrease in pain. There was no history of fever, systemic complaints, or any spinal or knee injury preceding the disease. His mother had received treatment for pulmonary tuberculosis 4 years previously. Dermatologic examination revealed well‐defined, brownish papules, plaques, and nodules of variable sizes, ranging from 3 cm to 6 cm, present over the right knee and front of the thigh in a linear pattern. The lesions were soft in consistency with ‘‘apple‐jelly’' nodules and the surrounding skin showed atrophic scarring (Fig. 1). 1 Plaques and nodules in a sporotrichoid distribution over the right lower limb (Case 1) All the hematologic and biochemical investigations were normal except for an elevated erythrocyte sedimentation rate (ESR) (48 mm/1 h). Venereal disease research laboratory (VDRL) test, enzyme‐linked immunoabsorbent assay (ELISA) for human immunodeficiency virus (HIV), and sputum examination for acid‐fast bacilli (AFB) were negative. Mantoux test with 1 tuberculin unit (TU) of purified protein derivative‐standard (PPD‐S) showed an induration of 25 × 20 mm. Blood culture and culture of the tissue homogenate for Mycobacterium tuberculosis, atypical mycobacteria, and fungal organisms revealed no growth. Slit skin smears from the plaque showed only lymphocytes and a few neutrophils. Immunoenzymatic test (ELISA) for antimycobacterial immunoglobulin M (IgM) antibody against the A 60 antigen complex was strongly positive (> 1 : 200 U). Histopathologic examination revealed a hypertrophic epidermis with noncaseating tuberculoid granulomas consisting of lymphohistiocytes, epithelioid cells, and Langhans' giant cells in the papillary and upper reticular dermis (Fig. 2a,b). No AFB were detected. Polymerase chain reaction (PCR) and guinea pig inoculation could not be performed due to a lack of facilities. 2 (a) Photomicrograph showing noncaseous tuberculoid granulomas in the papillary dermis (hematoxylin and eosin, × 40); (b) photomicrograph showing Langhans' giant cells (arrow) in the dense granuloma Skiagram of the chest was normal, although X‐ray of the lumbosacral spine revealed bony destruction of the sacral foramina. A magnetic resonance image (MRI) in the sagittal and axial plane was suggestive of caries of the fifth lumbar and sacral vertebrae with a large presacral and anterior epidural collection (Fig. 3). Computed tomography (CT) scan‐guided abscess drainage of the sacral region was performed and microscopic examination of the fluid revealed lymphocytes, polymorphs, macrophages, and a few AFB in a necrotic background. 3 Magnetic resonance image in the sagittal plane: caries ...

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Evaluation of acridinium-ester-labeled DNA probes for identification of Mycobacterium tuberculosis and Mycobacterium avium-Mycobacterium intracellulare complex in culture

Cited by 118 publications

References 15 publications

Evaluation of the semiautomated Abbott LCx Mycobacterium tuberculosis assay for direct detection of Mycobacterium tuberculosis in respiratory specimens

Evaluation of the semiautomated Abbott LCx Mycobacterium tuberculosis assay for direct detection of Mycobacterium tuberculosis in respiratory specimens

PCR assay based on DNA coding for 16S rRNA for detection and identification of mycobacteria in clinical samples

An unusual episode of lupus vulgaris masquerading as sporotrichosis

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