[Clinical study on the cases in which INH or RFP was discontinued during treatment for pulmonary tuberculosis].

Short course regimens; 2HRZ (E)(S)/4HR (E), 6HRS (E)/3-6HR and 6-9HR have been accepted as a standard chemotherapy (SC) for initial treatment of pulmonary tuberculosis in Japan. We studied the frequency of the treatment completion, the causes of the treatment failure and the outcome of the patients in whom INH or RFP was discontinued within 6 months after starting SC. The subjects included 597 newly diagnosed culture positive pulmonary tuberculosis patients admitted to 16 national hospital in 1996. Results were as follows. 1. In 47 (7.9%) of the 597 patients, either INH (19; 3.2%) or RFP (33; 5.5%) was discontinued. These 47 cases were defined as a SC incompleted group and the other 550 as a SC completed group. 2. The patients in the SC incompleted group were seen more frequently in the ages of 20s (11.9%), 50s (10.9%), 60s (11.7%) or 70s (11.4%). 21 (13.6%) of 154 female patients and 26 (5.9%) of 443 male patients were in the SC incompleted group. 3. The causes of cessation of INH or RFP were drug side effects (33; 5.5%), drug resistance (10; 1.7%) and complications or underlying diseases (8; 1.3%). 4. Fever or eruption (19; 3.2%) and drug induced hepatitis (12; 2.0%) were frequently seen as drug related side effects causing the cessation of INH or RFP. 5. The rate of culture negative conversion of TB bacilli at 6 months after the start of the treatment was 98.9% in the SC completed and 88.9% in the SC incompleted group respectively. In the SC incompleted group, there were three cases continuously positive and two other patients who relapsed and became culture positive again. In these five patients, INH or RFP was discontinued because of drug resistance.

[A case of drug-resistant pulmonary tuberculosis treated successfully following disappearance of rifampicin resistance after 17 years' chemotherapy].

A female who first acquired pulmonary tuberculosis in 1962 when she was 25 years old, admitted to the National Hiroshima Hospital in 1982. Her sputum has been smear positive for acid-fast bacilli for 3 years before admission in spite of continuous antituberculous chemotherapy, and were resistant to isoniazid (INH) and rifampicin (RFP). She was treated with a regimen containing ethambutol (EB), prothionamide (TH) and enviomycin (EVM) but continued to be culture positive. Though she was treated with various regimens which include one to three sensitive drugs, her sputum continued to be positive for M. tuberculosis in the following 14 years. During the course, resistance to EB, TH, cycloserine (CS) and streptomycin (SM) emerged. Resistance to RFP temporarily retracted in 1988, but her sputum was bacilli negative only for 2 months after the addition of RFP to previous regimen, and followed by resurgence of RFP resistance. In 1992, data of drug sensitivity tests showed sensitivity to TH, CS and RFP in turn, which were not used for 3 to 5 years. In 1993, she was treated with RFP, TH and EVM successfully and continued to be bacteriologically negative for 7 years so far. Drug resistance to M. tuberculosis is induced by inappropriate chemotherapy as seen in this case. Regimens with less than three drugs without RFP and INH was not only insufficient to get cure but, what was worse, also induced additional resistance to used drugs. The reason of successful chemotherapy in this case was spontaneous disappearance of drug resistance to RFP and TH. This case suggests that the disappearance of drug resistance is possible, when drugs are not used for more than a few years, hence the successful treatment could be expected. However it must be emphasized that the drug resistance is produced by incorrect treatment as seen in this case, and its prevention is of the prime importance.

[Desensitization therapy for antituberculous drugs].

We retrospectively evaluated the effectiveness of desensitization therapy for antituberculous drugs (Rifampicin and Isoniazid) in 28 cases (29 episodes) with adverse reactions to these drugs. Desensitization therapy for RFP was performed in 23 cases (24 episodes) with administration of a first dose of 1-150 mg and a final dose of 300-450 mg for 1-29 days. The success rate of this therapy was 79% (19 of 24 episodes). Desensitization therapy for INH was performed in 12 cases with administration of a first dose of 2.5-100 mg and a final dose of 200-400 mg for 3-25 days. The success rate of this therapy was 83% (10 of 12 cases). Based on a comparative study of cases between successful and unsuccessful desensitization to RFP and INH it was concluded that there were no significant differences with regard to allergic history, adverse effects and their periods of appearance, the first dose and final dose of administration and the interval of administration, starting periods of the desensitization therapy and the periods of appearance of adverse effects due to this therapy. We evaluated desensitization therapy for two antituberculous drugs (RFP and INH) for tuberculous patients for whom the use of such drugs was restricted because of adverse effects, and we found it is a useful treatment, showing a high rate of success (80%).

[Two-step tuberculin skin test in nurse students and hospital employees].

Booster phenomenon (recall effect) of tuberculin skin test, which disturbs diagnosis of tuberculous infection, is prevalent among BCG vaccinated population. We retested 34 nurse students whose initial tuberculin reaction was smaller than 30 mm by erythema (Group A) and 53 hospital employees whose initial reaction was smaller than 20 mm by erythema (Group B). Among the people whose diameter of erythema was less than 10 mm by the first test, 88 percent (8/9) of group A and 43% (6/14) of group B showed reaction 10 mm < or = by erythema and among those whose induration was < 10 mm, 54% (6/11) of group A and 48% (12/25) of group B showed reaction 10 mm < or = in the second testing. Mean and standard deviation of [the difference between the diameter of the 2nd and the 1st testing] was +7.3 +/- 11.8 mm in group A, +9.8 +/- 11.1 mm in group B by erythema and +2.6 +/- 5.9 mm in group A, +2.9 +/- 5.1 mm in group B by induration. These results indicate that booster phenomenon is highly prevalent among the tested group and there can be no appropriate criteria to distinguish new infection and booster phenomenon. Though two-step tuberculin skin test is recommended to get rid of booster phenomenon. Only a little is known about the value of this test to diagnose new infection in Japanese population, majority of whom being BCG vaccinated. Further investigations are required to apply two-step tuberculin skin test for diagnosis of new infection among hospital employees and health care workers in Japan.

Sternal tuberculosis in a 9-month-old infant after BCG vaccination.

UNLABELLED: Tuberculosis osteitis is mainly observed as a late complication of the pulmonary infection. We describe a 9-mo-old Japanese infant who became infected with Mycobacterium tuberculosis even after receiving Bacillus Calmette-Guerin (BCG) vaccination at 3 mo of age. This is the first report that the sternum is the localization for tuberculous osteitis in infants. He developed a localized tumor in the sternum without any respiratory symptom. Mycobacterium tuberculosis complex was detected by means of nucleic acid amplification test. Gastric aspirates also yielded Mycobacterium tuberculosis. The source of the infection was unclear. CONCLUSION: Tuberculous osteitis should be excluded in infants with undiagnosed bone lesions, even if they have been vaccinated with BCG.

[Desensitization therapy for allergic reactions of antituberculous drugs--evaluation of desensitization therapy according to the guideline of the Japanese Society for Tuberculosis].

We prospectively evaluated the effectiveness of desensitization therapy for cases showing side-effects to antituberculous drugs (Isoniazid and Rifampicin) according to the guideline proposed by the Treatment Committee of the Japanese Society for Tuberculosis. Nineteen patients (23-88 years old, male 9, female 10) who had experienced adverse effects after receiving antituberculous drugs and underwent desensitization therapy between August 1998 and March 2000 were studied. Underlying diseases were 14 cases of pulmonary tuberculosis, 2 cases of cervical tuberculous lymphadenitis, 1 case of pulmonary atypical mycobacteriosis, 1 case of pulmonary tuberculosis and tuberculous pleuritis, 1 case of pulmonary tuberculosis and tuberculous lymphadenitis. The regimens of treatment for tuberculosis were INH + RFP + EB in 8 cases, INH + RFP + EB + PZA in 7 cases, INH + RFP + SM in 2 cases, INH + RFP + SM + PZA in 1 case, and INH + RFP in 1 case. Adverse reactions were 8 cases of eruption, 7 cases of drug fever, 3 cases of drug fever and eruption, and 1 case of drug fever and cervical lymphadenopathy. The causative drugs suggested from DLST or the clinical course were RFP in 17 cases and INH in 8 cases. The clinical effect of desensitization therapy for these antituberculous drugs was good in 14 out of the 17 cases (82%) for RFP, and in 6 out of 8 cases (75%) for INH. The effectiveness rate of the present desensitization therapy according to the guideline of the Japanese Society for Tuberculosis was almost equal to that of previous desensitization therapy, and the clinical results were almost same in present and previous studies despite the different methods of administration of the antituberculous drugs.

[Tuberculin skin testing in students of school of nursing].

Tuberculin skin testing in students of the School of Nursing Attached to National Hiroshima Hospital was analyzed. On initial test using 0.05 microgram of PPDs, diameter of erythema in 26.7% of 300 new students were less than 9 mm. Twelve of 24 who were tested by two-step method reacted more than 10 mm on the second test. Twenty-seven non-reactors who were vaccinated with BCG all reacted more than 10 mm after 9 to 16 weeks after vaccination. They might be vaccinated in the past by insufficient technique and better be revaccinated. Thirty-one of 49 students who graduated in 1998 were tested and their reactions were compared with those on entrance or after BCG vaccination. The two tests were spaced 31 to 34 months apart. The reactions were weakened in the cases after BCG vaccination, and in those who were not vaccinated on entrance, only a little booster effect were observed, except in 3 graduates whose reactions were significantly boosted and thought to be infected while in school. As there is considerable variation in tuberculin reactivity after BCG vaccination, diameter of reaction should be kept on personal health record as base line reaction to diagnose tuberculous infection henceforce.

Pathological and radiological changes in resected lung specimens in Mycobacterium avium intracellulare complex disease.

The present study was designed to evaluate the pathological and immunohistochemical findings of Mycobacterium avium intracellulare complex (MAC) lung infection. A retrospective study was performed in five cases with positive cultures for MAC in whom lung resections were performed between January 1989 and December 1996. A determination of whether or not MAC caused pulmonary disease was made using the 1997 criteria defined by the American Thoracic Society. In addition, MAC was cultured from all of the five lung specimens. Pathological and immunohistochemical findings as well as chest computed tomography (CT) findings were evaluated in these five patients. Pathological findings of bronchiectasis, bronchiolitis, centrilobular lesion, consolidation, cavity wall and nodules were demonstrated, respectively, in relation to chest CT findings. Extensive granuloma formation throughout the airways was clearly demonstrated. Immunohistochemical staining demonstrated: 1) epithelioid cells and giant cells; 2) myofibroblasts extensively infiltrating the cavity wall; and 3) B-cells detected in aggregates in the vicinity of the epithelioid granulomas. This study identified pathological and immunohistochemical characteristics of Mycobacterium avium complex infection relative to chest computed tomography findings and allowed the conclusion that bronchiectasis and bronchiolitis were definitely caused by Mycobacterium avium complex infection.

[Nontuberculous mycobacteriosis; the present status and in the future. 3-(2) Long-term prognosis of Mycobacterium avium complex disease].

This paper shows long-term course of 71 patients with Mycobacterium avium complex (MAC) disease treated in National Hiroshima Hospital between 1977 and 1991 and were observed for more than 5 years or died between 6 months and years from the onset. In the patients who were not treated with chemotherapeutic agent or treated with less than 3 drugs, about 60 per cent deteriorated and few obtained persistent negativity of bacteria. In patients treated with at least 3 drugs out of isoniazid, rifampicin, ethambutol, ethionamide, cycloserine, clarithromycin and new-quinolones for more than 12 months, 59.2 per cent deteriorated, while 29.6 per cent obtained negative conversion of bacteria by initial chemotherapy. However only 18.5 per cent remained negative during long term observation. In patients treated with at least 3 drugs including one aminoglycoside such as streptomycin for more than 12 months, 64.7 per cent obtained negative conversion and 35.5 per cent remained negative bacteriologically, while 23.5 per cent deteriorated. In twelve patients who were operated, only one patient deteriorated. The long-term prognosis of MAC disease under the chemotherapy mainly by antituberculous agents up to this time was still poor compared to that by short-term observation. Anticipated new regimens for MAC should be multidrug, to prevent secondary drug resistance.

[An observation on tuberculosis associated with HIV infection in Japan].

OBJECTIVE: To observe the reported cases of tuberculosis (TB) with HIV infection in Japan, in terms of their main clinical features and related factors. METHODS: A voluntary reporting network has been organized by the authors who are specialists of TB or respiratory medicine in tuberculosis institutions located roughly all over the country. The members have been encouraged to report not only their own cases but cases seen by their friends or in other institutions. RESULTS: By the end of 1996, a total of 71 cases have been reported of which 59 were TB and 12 NTM cases. Nationality of the cases were; Japan 48, Other Asian countries 16, Others 7. All of the NTM cases were Japanese. 30% of the cases were aged less than 30 years, 24% were thirties, 24% forties, 17% fifties and 6% were those aged 60 years or older. The cases were clearly younger than the TB cases in the national TB registry, and older than HIV-infected persons as known from the HIV surveillance system. 97% of the TB cases were bacteriologically confirmed cases. Eight of NTM cases were positive for MAC, others for M.kansasii. 42% of the cases had extra-pulmonary disease, including disseminated infections seen among 19%. Of TB cases 25% were excreting bacilli resistant to any of the anti-TB drugs which was higher than in the case of general TB population (10-15%). 11% of TB cases had past history of TB treatment. The cases had severe immunological impairment, 79% of the cases having CD4+ cell count less than 100. The route of HIV infection were; 51% heterosexual, 13% homosexual, 13% through blood preparations, etc. DISCUSSION: Although there may be many cases not included in this observation, it is considered to well reflect the real situation of the problem of Japan. More attention should be paid to HIV infection of the patients in the clinical practice of TB in Japan.

[Tuberculosis found among foreigners and treated at the National Hiroshima Hospital or Hiroshima Anti-Tuberculosis Association].

In this study, we investigated 31 foreign patients who had been diagnosed as having pulmonary tuberculosis at the National Hiroshima Hospital or Hiroshima Anti-Tuberculosis Association between 1982 to 1995. Their age ranged from 17 to 63 years, and all of them had been in Japan for less than five years. Twelve of the patients came from South America, 18 from Asia, and one from Egypt. Seven Peruvians had been infected in a tuberculosis outbreak. The reasons of their immigration were job-seeking (14 out of 31), education (8 out of 31), and marriage to Japanese men (6 out of 31). None of them were illegal immigrants. In four women who had married Japanese men, various symptoms such as cough, sputum, and slight fever came on during pregnancy, however, none of them had been diagnosed as having tuberculosis, and consequently they had received no medication during pregnancy. In 6 out of the 8 patients who were college students, their sputa were negative for tubercle bacilli, and they could be treated by chemotherapy as outpatients. In provincial cities, every effort should be made to detect tuberculosis among members of the foreign community who are living under different environments and conditions.

[The epidemiology of nontuberculous mycobacterial diseases in Hiroshima Prefecture].

The patients of diseases caused by nontuberculous mycobacteria (NTM) newly diagnosed in 1993 in Hiroshima Prefecture were investigated retrospectively. During 1993, 59 newly diagnosed patients of NTM disease, who were the resident of Hiroshima Prefecture were reported from 6 hospitals. Mycobacterium avium complex (MAC) disease was most frequent (53 patients, 90%) and M. kansaii (MK) disease, which accounts for more than 20 percent of all the NTM diseases in the national survey of Japan, was found only in 3%. Other pathogenesis were M. chelonae and M. fortuitum. Incidence rate for all NTM disease per 10(5) population was 2.1. Age-specific incidence rate calculated from the estimated population of Hiroshima Prefecture was; less than 0.4/10(5) under the age 40, 4.0/10(5) in the age group 50 to 59, 4.5/10(5) in the age group 60 to 69 and 9.4/10(5) in the age group over 70. Thirty-one patients (57%) had underlying pulmonary disease such as previous lung tuberculosis (TB) and previous history of TB was found in 41 percent of patients over 60 yr. of age. The data suggest that epidemiological picture of NTM diseases may change in the future. Relative increase of elder population will lead to the increase of NTM diseases as a whole; the incidence rate calculated from the age specific incidence rate and predicted population of Japan in 2025 is 3.1/10(5), which is 1.5 fold higher than the present rate. However, secondary NTM diseases with healed TB will decrease because of the sharp decline of TB morbidity in Japan. Assuming that secondary NTM disease with healed TB decreases to one-fourth, estimated incidence rate will be 2.3. Further, the upward tend of MK disease in Japan should be taken into consideration. Thus, the future trends of NTM diseases will increase fairly due to the increase of elder population as well as the MK disease. It was pointed out that many NTM disease cases had been counted as tuberculosis in the surveillance system of Japan. In our present study, 52 (82%) were registered as tuberculosis in the beginning and only 19 cases were reported later to health centers as NTM disease. As tuberculosis classification and surveillance system in Japan were changed from 1996 to separate NTM disease from TB, NTM disease cases will be counted separately in the tuberculosis statistics. This revision in the management of NTM disease should be taken into account when discussing the trend of tuberculosis statistics in Japan.

[Drug susceptibility test for Mycobacterium tuberculosis using non-radioactive substance, alpha-antigen].

BACTEC system is a reliable and rapid drug susceptibility test for mycobacteria and is widely accepted in Europe and in the United States of America. In Japan, it is impossible to introduce the BACTEC system in clinical laboratories because of strict regulations for the use of radioactive substances in Japan. To resolve this dilemma, we adopted alpha-antigen (alpha-antigen), a widely distributed secretory protein of mycobacteria, as the index substance replacing the radioactive substance 14C in BACTEC system. Alpha-antigen was detected by reverse passive latex agglutination (LA), using latex sensitized with rabbit anti-alpha-IgG. Susceptibility of 17 M. tuberculosis strains isolated from patients, grown on Ogawa egg medium and then cultured in 7H9 medium, and of 46 M. tuberculosis strains freshly isolated from patients by 7H9 medium of MB check system, was tested for four antituberculosis drugs, isoniazid (INH), rifampicin (RFP), streptomycin (SM) and ethambutol (EB). Ninety four percent of the control cultures were positive for alpha-antigen within 7 days after the inoculation. The MIC values of H37Rv strain in 7H9 medium determined by the method of Heifets were 0.05 micrograms/ml for INH, 0.03 micrograms/ml for RFP, 0.025 micrograms/ml for SM and 1.9 micrograms/ml for EB. In 17 strains from Ogawa egg medium, the results obtained from all but 4 strains for SM, 1 for INH, 1 for RFP and 7 for EB were concurrent with that obtained by the method using 1% Ogawa egg medium. No strains were determined to be resistant to any drug by the alpha-antigen method and be sensitive by the Ogawa medium method. In 46 strains cultured by the MB check system, the results of 42 strains for SM, 35 for INH, 39 for RFP and 36 for EB coincided with those determined by the Microtiter method. Among the strains determined to be resistant by Microtiter method, 1/2 for SM, 10/14 for INH, 4/17 for RFP and 8/10 for EB were determined to be sensitive by alpha-antigen LA method. The disagreement was seen mostly in strains which were determined to be resistant by the method using egg medium, while sensitive by the alpha-antigen LA method. The discrepancy might originate from the difference of critical concentration due to heat inactivation of the drugs and absorption in the egg medium. However, some instability was observed in latex agglutination and its cause should be examined further. This method of utilizing 7H9 medium for culture and alpha-antigen as the index of mycobacterial growth can be an expedient and economical drug susceptibility test because it does not use radioactive substance as in the case of BACTEC system.

Secretion of MPB64 antigen by a recombinant clone of Mycobacterium smegmatis: characterization and application for the diagnosis of tuberculosis.

MPB64, a specific antigen to Mycobacterium tuberculosis complex (TB complex), was produced and secreted by a clone of M. smegmatis-MPB64 where the structural gene of MPB64 was inserted using a new mycobacteria, E. coli shuttle plasmid pNIS vector. Antibodies against the recombinant MPB64 (rMPB64) were used for the reverse particle latex agglutination (RPLA) test to detect the MPB64 antigen rapidly. RPLA tests were applied to the shock cultures and the clinical isolates of mycobacteria to identify TB complex. RPLA with anti-MPB64 antibody-coated latex beads completely distinguished TB complex from other mycobacteria. Thus, it is suggested that RPLA with anti-MPB64 antibody would be a new, easy and inexpensive method for the rapid diagnosis of tuberculosis.

[Management of nontuberculous mycobacteriosis in health centers and tuberculosis surveillance system].

Nontuberculous Mycobacteriosis (NTM) is a different disease from tuberculosis, but in Japan most of them are still obliged to be treated as tuberculosis under medical insurance scheme and included statistically in the number of tuberculosis. In this investigation, patients of NTM who were at first registered as tuberculosis and diagnosed later as NTM or found to be positive for nontuberculous mycobacteria were analysed from the standpoint of tuberculosis statistics and activities at health centers. Out of 1207 newly registered tuberculosis in 1993 at 23 health centers (HCs) or its branches in Hiroshima Prefecture, 482 cases were bacteriologically positive, and among them 40 cases were found to be NTM later. Under the current tuberculosis surveillance system, 4 cases from 1 HC were omitted from the tuberculosis registry, 10 cases from 4 HCs were kept on the tuberculosis registry as 'culture positive for nontuberculous mycobacterium', 15 cases from 7 HCs were registered as 'tuberculosis with nontuberculous mycobacteriosis as a complication' and in 11 cases no informations on NTM were entered into the registry. Only 6 smear positive cases which were registered as 'culture positive for nontuberculous mycobacteria' were excluded from the number of smear positive tuberculosis under the surveillance system. In other investigation made by hospitals in the same area on NTM, 59 patients with definite NTM were reported in 1993. At least 52 were registered first as tuberculosis, thus NTM occupies at least 4.3 percent of all newly reported tuberculosis and 10.8 percent of new smear positive cases.(ABSTRACT TRUNCATED AT 250 WORDS)

[An outbreak of tuberculosis involving foreign workers from South America].

A twenty-four year old male Peruvian of Japanese origin, who came to Japan in September 1990 and had been working in a minor factory in a rural area, was admitted to a hospital in March '91 with severe cough. Smear examination of his sputum smear was positive for acid-fast bacilli and his chest X-ray showed multiple cavities (Index case). Subsequent contact examination identified further four patients with pulmonary tuberculosis among his colleagues in the factory, all of whom lived in the same house with the index case. During following three years, further six patients with mycobacteriosis, two Peruvians and four Japanese, were found among the employee of that factory. M. tuberculosis was cultured from the sputa obtained from seven of these eleven patients. Another patient was diagnosed as non-tuberculous mycobacteriosis. Restriction fragment length polymorphism (RFLP) analysis carried out with five strains of M. tuberculosis isolated from these patients revealed the identical RFLP pattern which is uncommon in Japan. Still more, an isolate from another patient was subjected to RFLP analysis by chance, and was found to show the same RFLP pattern. Later epidemiological study revealed that the last patient, a 53 year-old saleswoman of boxlunch, might have some contact with the index case at her booth. Though RFLP analysis was not done for the isolate from the index case, from the identity of RFLP patterns of other isolates, clinical course and epidemiological study, it is considered that six patients were certainly, and two others were probably infected from the index case.(ABSTRACT TRUNCATED AT 250 WORDS)

[Tuberculin sensitivity to purified protein derivatives from Mycobacterium other than tuberculosis (PPD-B, PPD-Y, PPD-F and PPD-C) and PPDs among patients with mycobacteriosis--cooperative study of the Research Committee for the Mycobacteriosis in Japan].

Purified protein derivatives (PPDs) prepared from M. intracellulare (PPD-B), M. kansasii (PPD-Y), M. fortuitum (PPD-F), M. chelonei subsp. abscessus (PPD-C) and M. tuberculosis (PPDs) were simultaneously used in skin tests on patients diagnosed as having tuberculosis or atypical mycobacteriosis to reveal their specificity, clinical usefulness and immunological status of the patients. The mean diameter of reaction (redness) for patients with M. tuberculosis positive sputum (TB group, n = 71; age, 20-90 yrs) was PPDs, 20.4 mm; PPD-B, 7.9 mm; PPD-Y, 11.7 mm; PPD-F, 0.8 mm; and PPD-C, 0.3 mm. For M. avium complex positive patients (MAC group, n = 100; age, 31-89 yrs), the results were PPDs, 10.9; PPD-B, 16.9 mm; PPD-Y, 10.7 mm; PPD-F, 1.6 mm; and PPD-C, 0.3 mm. The M. kansasii positive patients (K group; n = 8) showed results of PPDs, 12.6 mm; PPD-B, 10.7 mm; PPD-Y, 20.8 mm; PPD-F, 0.5 mm; PPD-C, 0.0 mm. The M. fortuitum positive patients (F group; n = 5) had measurements of PPDs, 5.8 mm; PPD-B, 4.4 mm; PPD-Y, 9.8 mm; PPD-F, 17.8 mm; and PPD-C, 16.0 mm. The patients who were previously M. tbc. positive but presently negative patients (pre. TB group; n = 50) showed the following results: PPDs, 16.6 mm; PPD-B, 7.4 mm; and PPD-Y, 10.9 mm. For the patients who were previously M. avium complex positive (previous MAC group; n = 19), the results were PPDs, 10.4 mm; PPD-B, 9.9 mm; and PPD-Y, 7.7 mm. Also considering their frequency distribution curve, with exception of the previous MAC group, the patient groups showed specificity to the PPD of the bacilli detected. The previous MAC group recorded no significant difference in response to PPDs and PPD-B. Strong cross reactions were observed between PPD-F and PPD-C, and moderate reactions between PPDs, PPD-B and PPD-Y. Cross reactions were scarce between PPDs, PPD-B or PPD-Y and PPD-F or PPD-C. Though it is difficult to distinguish cross-reaction and multiple infections, majority of the patients (72-85%) showed greatest response to the PPD that corresponds with the species of bacilli detected. In conclusion, two or more PPDs applied simultaneously can be of aid in diagnosing mycobacteriosis especially in the early stages of the disease. Also, cross-reactions between atypical mycobacteria and PPDs should be taken into consideration when diagnosing infection caused by M. tuberculosis.

[Tuberculin sensitivity to purified protein derivatives (PPD) from M. intracellulare (PPD-B), M. kansasii (PPD-Y), M. fortuitum (PPD-Y) and M. tuberculosis (PPDs) among healthy volunteers].

To reveal the epidemiology of mycobacteria other than Mycobacterium tuberculosis in Japan, we tested 379 healthy volunteers from Self Defence Force Army stationed in Hiroshima Prefecture with 0.05mcg of PPDs, 0.1mcg of PPD-B, PPD-Y and PPD-F. Majority of the volunteers had been immunized with BCG by thirteen years old. Rate of positive reaction (diameter of redness > or = 10mm) in each PPD in each age group were; PPDs [18-19yr (n = 39); 30.8%, 20-29yr (n = 178); 63.5%, 30-39yr (n = 78); 91.0%, 40-53yr (n = 84); 92.9%], PPD-B [12.8%, 24.7%, 38.5%, 48.8%], PPD-Y [5.1%, 14.6%, 26.9%, 26.2%], PPD-F [0%, 10.1%, 12.8%, 10.7%]. Frequency distribution curve of PPDs in age groups above 20 years old had a peak at about 14mm of diameter, while low-responder were dominant in age 18 to 19. In PPD-B, there were two peaks, one in less than 5mm and the other between 10 to 15mm which was considered as the group sensitized by M. avium complex and became larger in older age group. In PPD-Y and in PPD-F, the frequency distribution showed an exponential curve with a little shift to right in older group in PPD-Y. As there was a considerable degree of cross-sensitivity, we provisionally regarded the maximum reaction of four PPD (larger than 5mm) or the reaction which is 75 per cent or more of each person's maximum reaction as specific.(ABSTRACT TRUNCATED AT 250 WORDS)

[Rapid diagnosis of mycobacteria, especially Mycobacterium tuberculosis and Mycobacterium avium complex, using MB Check system].

Fourty-five sputum specimens were subjected to isolation for mycobacteria either MB Check system (MB method; F. Hoffmann-La Roche Ltd., Basel, Switzerland) or 3% Ogawa egg medium (Ogawa method). Test sputum was treated with 4 volumes of 4% NaOH for 1-2 min and 0.1 ml of the resulting mixture was inoculated onto 3% Ogawa egg medium. The remaining portion of the mixture was neutralized with IN HCl, diluted with 1/15 M phosphate buffer (PB; pH 6.8), and subsequently centrifuged at 3,000 rpm for 20 min. The sediment was suspended in 1.5 ml of PB and 0.5 ml each was inoculated into MB Check M bottle (20 ml) supplemented with M supplement (1 ml). In MB method, bacterial growth was measured on Middlebrook 7H11 agar medium and Middlebrook 7H11 agar medium containing NAP (p-nitro-alpha-acetylamino-beta-hydroxy-propiophenone). Among 45 sputum specimens, the number of positive specimens for mycobacterial growth in the above two cultivation methods and time required for growth were as follows: 3% Ogawa egg medium; 12 specimens (26.7%) gave positive growth, including 7 of M. tuberculosis complex strains on 14-35 days (average 22 days) and 5 of M. avium complex strains on 14-21 days (average 18 days); MB method; 15 of specimens (33.3%) gave positive growth, including 8 of M. tuberculosis complex strains on 7-21 days (average 15 days), 6 of M. avium complex strains on 7-14 days (average 11 days) and 1 of M. scrofulaceum strain on 28 days. There was no specimen which was positive for mycobacterial growth on 3% Ogawa egg medium but negative on MB medium.

[Rapid diagnosis of mycobacteria, especially Mycobacterium tuberculosis and Mycobacterium avium complex, using BACTEC 460 TB system].

Fourty-five sputum specimens collected at the National Sanatorium Hiroshima Hospital were subjected to cultivation using either BACTEC 460 TB System (BACTEC method; Becton Dickinson Co., Towson, Md., U.S.A.) or 3% Ogawa egg medium. Test sputum was treated with four volumes of 4% NaOH for approximately two minutes, after which 0.1 ml of the treated sputum was immediately inoculated onto 3% Ogawa egg medium. After neutralizing the remaining pretreated sputum with 1N HCl, and diluting with 1/15 M phosphate buffer PB; pH 6.8), it was then centrifuged at 3,000 rpm for 20 min and the sediment was suspended in 1.5 ml of PB. Volumes of 0.5 ml each were inoculated into BACTEC 12B medium (4 ml), containing PANTA for prevention of contamination and POES for promoting the growth of mycobacteria. In the BBCTEC method, bacterial growth was measured in terms of increases in the Growth Index (GI) values which were determined by the amount of 14CO2 released from the 14C-labelled palmitate during cultivation at 37 degrees C (positive growth; GI greater than or equal to 50). Moreover, rho-nitro-alpha-acetylamino-beta-hydroxy-propiophenone (NAP)-sensitivity testing was done by transferring a part of the BACTEC 12B culture showing positive growth to a NAP vial, and thereafter subjected to further cultivation.(ABSTRACT TRUNCATED AT 250 WORDS)