Dry-reagent-based PCR as a novel tool for laboratory confirmation of clinically diagnosed Mycobacterium ulcerans-associated disease in areas in the tropics where M. ulcerans is endemic.

After tuberculosis and leprosy, Buruli ulcer (BU), caused by Mycobacterium ulcerans, is the third most common mycobacterial disease in immunocompetent humans. The disease occurs in tropical countries, with foci in West Africa, Central Africa, and the western Pacific. BU is defined as an infectious disease involving the skin and the subcutaneous adipose tissue characterized by a painless nodule, papule, plaque, or edema, evolving into a painless ulcer with undermined edges and often leading to invalidating sequelae. Due to the fundamental lack of understanding of modes of transmission, disease control in endemic countries is limited to early case detection through improved active surveillance and surgical treatment. The laboratory confirmation of BU is complicated by the absence of a diagnostic "gold standard." Therefore, misclassification and delayed diagnosis of BU may occur frequently, causing a considerable socioeconomic impact in terms of treatment costs due to prolonged hospitalization. In order to respond to the urgent need to develop reliable tools for early case detection and to overcome technical difficulties accompanying the implementation of diagnostic PCR procedures in tropical countries, a dry-reagent-based PCR formulation for the detection of M. ulcerans in diagnostic specimens has been developed at the Bernhard Nocht Institute for Tropical Medicine. Following technical and clinical validation, the assay has been successfully installed and field tested at the Kumasi Centre for Collaborative Research in Tropical Medicine, Kumasi, Ghana. Preliminary results show an excellent diagnostic sensitivity of >95%.

Use of a standardised checklist to assess peripheral sputum smear microscopy laboratories for tuberculosis diagnosis in Uganda.

SETTING: TB diagnostic units in Uganda. OBJECTIVES: To assess and improve the supervision and performance of sputum smear microscopy in the peripheral diagnostic units in Uganda using a standardised laboratory checklist. DESIGN: A standardised checklist was developed and used during the quarterly supervisory visits of the District TB and Leprosy Supervisors for five quarters from the fourth quarter of 1997 until the last quarter of 1998. Individual peripheral laboratory performance was monitored during the study period. RESULTS: Forty-eight of 304 TB diagnostic units in six of the 45 Ugandan Districts were supervised using the checklist. A total of 208 checklists were analysed. The situational analysis of the peripheral diagnostic units at the beginning and at the end of the study showed a marked improvement in laboratory performance in all aspects related to sputum smear microscopy. Individual laboratory performance was monitored over five quarters, and timely response to shortcomings was provided. CONCLUSION: The systematic use of a standardised laboratory checklist can be considered an important step forward in improving the performance of the peripheral laboratories in Uganda through on-the-spot correction of any identified shortcomings.

Anti-tuberculosis drug resistance surveillance in Uganda 1996-1997.

SETTING: Drug resistance surveillance conducted by the National Tuberculosis and Leprosy Control Programme (NTLP) Uganda from 1996-1997 in collaboration with the Armauer Hansen Institute/German Leprosy Relief Association (GLRA), Germany, for the WHO/IUATLD Global Project on Anti-Tuberculosis Drug Resistance Surveillance. OBJECTIVE: To determine the prevalence of primary and acquired anti-tuberculosis drug resistance in Uganda. DESIGN: The survey area covered three GLRA-supported operational NTLP zones, corresponding to 50% of the Ugandan population. A representative random sampling of individual patients was chosen as sampling procedure. Altogether 586 smear-positive TB patients (537 new cases and 49 previously treated cases) were included in the survey. RESULTS: For primary resistance the results were as follows: isoniazid (H) 6.7%, rifampicin (R) 0.8%, ethambutol (E) 6.1%, streptomycin (S) 13.4%, thioacetazone (T) 3.2%, pyrazinamide (Z) 0%, multidrug resistance (MDR) 0.5%; for acquired resistance they were: H 37.8%, R 4.4%, S 22.2%, E 11.1%, T 20.0%, Z 0%, and MDR 4.4%. CONCLUSION: According to these data the NTLP Uganda has been effective in preventing high levels of primary drug resistance. If it is assumed that the sampling process reflects the distribution of new patients and previously treated patients in the study areas, the amount of acquired resistance (any resistance) in the community of smear-positive patients is approximately 5%. To further monitor programme performance the NTLP will embark on a nationwide survey in 1998/1999.

Tuberculosis chemotherapy and sputum conversion among HIV-seropositive and HIV-seronegative patients in south-eastern Uganda.

OBJECTIVE: To investigate if there is a difference in response to tuberculosis treatment between HIV seronegative and HIV seropositive patients following two months of intensive phase tuberculosis treatment. DESIGN: Prospective cohort study. SETTING: St. Francis Leprosy Centre, south-east Uganda. SUBJECTS: Four hundred fifty seven patients with never previously treated sputum smear-positive tuberculosis admitted during a two-year period in 1991/1993. INTERVENTION: Intensive phase treatment with streptomycin, isoniazid, rifampicin and pyrazinamide. MAIN OUTCOME MEASURES: Sputum conversion from a positive to a negative smear at eight weeks of treatment. RESULTS: HIV seropositivity prevalence was 28%. Among HIV seronegative patients, conversion to a negative smear status occurred in 76% persons compared to 78% in HIV seropositive patients. This difference was not statistically significant (OR = 0.9; 95% CI, 0.6-1.5). HIV seropositive patients, however, were more likely to die (p = 0.017). A high prevalence of resistance to isoniazid and streptomycin was found. Isoniazid resistance was more likely in HIV seronegative patients with M. tuberculosis strains compared to HIV seropositive persons (p < 0.005). Initial resistance to antituberculosis drugs did not have a significant effect on smear conversion. CONCLUSION: This study demonstrates that HIV-seropositive status is not a principal factor in delaying sputum conversion among patients receiving intensive phase tuberculosis treatment.

PIP: A prospective cohort study was undertaken to investigate the response of HIV-seropositive and -seronegative patients at St. Francis Leprosy Center, southeastern Uganda, to tuberculosis chemotherapy. The study population included 457 patients without a history of prior tuberculosis therapy between 1991 and 1993. The subjects were exposed to an intensive phase therapy of rifampicin, streptomycin, isoniazid, and pyrazinamide. After the treatment, sputum culture and sensitivity tests were conducted. Findings showed that 77% of the patients who never received tuberculosis treatment in the past converted to a negative smear status after the 8-week treatment. There was no significant difference in sputum conversion rates between HIV-seropositive and -seronegative patients. The study also revealed that HIV seropositivity prevalence was 28%. Among HIV-seronegative patients, conversion to a negative smear status occurred in 76% compared to 78% HIV-seropositive patients. Moreover, a significant number of HIV-seronegative patients died during the initial course of the therapy. Also, a high prevalence of isoniazid and streptomycin resistance was noted; however, this result never affected the conversions of smears. In conclusion, the study clearly demonstrates that other factors outside the seropositive status may be the principal causes of the delay in sputum conversion among patients receiving intensive tuberculosis chemotherapy.

[Current aspects of leprosy]. / Aktuelle Aspekte der Lepra.

About 95% of individuals who come in contact with M. leprae do not develop overt disease. The clinical form of the disease correlates with the T-cell mediated immune response of the host rather the direct damage caused by bacilli. We review the current aspects of epidemiology, transmission, bacteriology, clinical features, reactions, diagnosis, chemotherapy and treatment of reactions.

[Leprosy--current aspects of a disease from biblical times]. / Lepra--aktuelle Aspekte einer Erkrankung aus biblischer Zeit.

95% of individuals who come in contact with M. leprae do not develop an overt disease. It begins as an indeterminate form that may undergo spontaneous cure or may progress to different forms of leprosy (TT, BT, unstable form of BB, BL, or LL). The clinical form of the disease correlates with the T cell mediated immune response rather than to the direct damage caused by the bacilli. The lack of cellular immunity in lepromatous patients relates specifically to M. leprae. Current aspects of etiology, transmission, epidemiology, classification, clinical features, immunopathology, chemotherapy, treatment of reactions, immunotherapy and vaccination are elucidated and discussed.