Neo-nocardin, a new antibiotic produced by a new species of Nocardia. II. The effect of neo-nocardin on the experimental tuberculosis in mice.
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The prevalence of pulmonary nocardiosis in a tuberculosis and chest diseases hospital in Amritsar is reported. Of 1510 sputum samples cultured from 1016 patients, 67 sputa originating from 16 patients were found to be positive for the Nocardia asteroides species complex. Based upon repeated isolation of N. asteroides from the respiratory tract, its microscopic demonstration in KOH wet mounts or stained smears of sputum and clinical evaluation of patients, 14 cases of pulmonary nocardiosis were diagnosed. This gave a prevalence of 1.4% pulmonary nocardiosis in the tuberculosis hospital. The prevalence of the disease was found to be 1.3% in the males as against 1.5% in the females. Of the various clinical categories of patients investigated, pulmonary tuberculosis with sputum negative for acid-fast bacilli (AFB) yielded the highest prevalence of 3.2%, followed by 1.3%, 1.2%, 1.1% and 0.5% in pneumonia, chronic obstructive pulmonary disease (COPD), bronchiectasis and pulmonary tuberculosis with sputum positive for AFB, respectively. Type IV cutaneous hypersensitivity to nocardin was observed in 19 of 908 (2%) patients tested, whereas only a solitary positive reactor was found among 260 healthy volunteers. Twelve of 19 nocardin positive reactors (63%) had unequivocally proven pulmonary nocardiosis. The nocardin skin test gave false negative results in two nocardiosis patients. More comprehensive investigations are warranted in order to evaluate the nocardin skin test as an additional aid for the diagnosis of nocardiosis. Barring a solitary exception, the nocardiosis patients were successfully treated with sulphadiazine or trimethoprim-sulphamethoxazole (TMP-SMZ) combination. To the best of our knowledge, this is the largest series of pulmonary nocardiosis patients in a prospective study as yet reported from India. The observations underscore the point that nocardiosis warrants greater attention in the differential diagnosis of bronchopulmonary diseases.
Concentrated cell-free filtrates (nocardins) were prepared from Nocardia asteroides cultures grown on Sauton's synthetic broth. Nocardins from 10 strains of six N. asteroides serotypes were produced and the proteins separated by isoelectric focusing. N. asteroides antigens among these proteins were tested for specificity using rabbit antisera and monoclonal antibodies (MAbs) against N. asteroides and Mycobacterium tuberculosis by the enzyme-linked immunoelectrotransfer blot test. At least 15 protein antigens were identified from each of the 10 nocardins. The immunodominant antigens were one serotype-specific N. asteroides protein with an isoelectric point (pI) of 4.0 (factor 1) and two group antigens with pIs of 4.43 (factor 6) and 4.68 (factor 8). The nitrocellulose strips prepared with these antigens did not react with antibodies to M. tuberculosis, nor with normal sera from humans, rabbits, or mice, but reacted specifically with anti-N. asteroides MAbs and polyclonal antibodies. Four purified protein derivatives of tuberculin were tested and did not cross-react with the three anti-N. asteroides MAbs. These reactions suggest that the antigens identified as factors 1, 6 and 7 are specific to N. asteroides and that factor 1 is specific for serotype 2, while factors 6 and 8 are species-specific.
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