In Japan the number of patient infected with M. avium complex (MAC) has been increasing in contrast to a decrease of pulmonary tuberculosis. DNA probe method enabled us be able to differentiate an isolated MAC into M. avium and M. intracellulare. From 1991 to 1992, we performed an investigation to apply this new technique of the DNA probe method on 52 patients of atypical mycobacteriosis diagnosed as infected with MAC by the ordinary method at the Higashi Nagoya National Hospital. The group consisted of 27 males and 25 females. M. avium infection was found in 39 patients (M. avium group) and M. intracellulare in 16 patients (M. intracellulare group). No significant gender difference was found between two groups. The M. avium group showed more complications in contrast to the M. intracellulare group. As complications in the M. avium group, pulmonary aspergillus infection, bacterial pneumonia and bronchiectasis were found in 4, 3 and 2 cases, respectively. The rate of drug resistance to antituberculosis drugs was high in the both groups. Chemotherapy with isoniazid (INH) rifampin (RFP) and streptomycin (SM) in five patients, that with INH, RFP and ethambutol (EB) in three were found to be effective after 4 months treatment. Three patients in M. avium group died of respiratory failure, asper gillus infection and renal failure. In contrast the prognosis of patients in the M. intracellulare group seemed to be better as there was no fatal case. We conclude that DNA probe method is useful to differentiate between M. avium and M. intracellulare, and enabled us to select more appropriate selection of the chemotherapy and to assess of the prognosis.

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The conventional drug susceptibility tests of Mycobacterium tuberculosis are time consuming and usually require 2 to 4 weeks to obtain final results. In this study, we attempted to develop a novel method for rapid detection of drug-resistant M. tuberculosis using hybridization protection assay (HPA).
Clinically isolated strains of M. tuberculosis including seven isoniazid (INH) susceptible strains and six resistant strains were used. The organisms grown on Ogawa egg medium were inoculated into Middlebrook 7H9 broth and cultured at 37°C for one week. Then, the inoculum of each strain was prepared as a tenfold dilution of bacterial suspension at McFarland No.0.5. The inocula were mixed with INH solutions to yield final concentrations of 0.1 and 1.0μEg/ml, and the resulting bacterial suspensions with or without test drug were cultured on the swaying plate at 37°C for up to 5 days. At intervals, 50μEl of each sample was withdrawn and subjected to the protocol of the HPA using acridinium-ester (AE) labeled DNA probe, and then the relative light unit (RLU) was read in a luminometer. In the case of the susceptible strains, a significant difference in the mode of increase in RLU ratio (% of RLU on day x/RLU on day 0) was observed between the INH-treated and drugfree control sets within three days of cultivation, while no such differences were seen in the case of the resistant strains.
Since this method was not only rapid, safe and simple, but gives the results well correlated to those by the conventional methods, it is expected that the method will be used widely in many laboratories.

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A 39 year-old male visited the hospital complaining of perianal pain, swelling and redness. Under the diagnosis of an anal abscess, drainage was performed repeatedly. As the wound failed to heal and fistulae were detected, excision of entire tract was performed. On histopathological examination of the resected fistulae, caseous necrosis, Langhans giant cells, and epithelioid cell infiltration were found and diagnosed as anal tuberculosis. Chest X-ray showed cavitary lesion with infiltrative shadow in right upper lobe. Acid-fast bacilli were positive in sputum, and the diagnosis of pulmonary tuberculosis was confirmed. Anti-tuberculosis therapy was immediately started with good response to treatment.
As tuberculosis of anal region is so rare recently and there is no characteristic clinical picture, it is very difficult to diagnose it pre-operatively. In some cases such as ours, pulmonary or other tuberculosis is accompanied with anal tuberculosis. Therefore, accurate diagnosis of anal tuberculosis is needed to find other tuberculosis early. As anal tuber culosis is rarely diagnosed correctly before operation on the basis of the clinical picture, the histopathological examination of the excised fistula is mandatory for the correct diagnosis of anal tuberculosis.

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