MODS Culture for Primary Diagnosis of Tuberculous Meningitis and HIV-Associated Pulmonary Tuberculosis in Indonesia.

Aims: To compare the microscopic observation drug susceptibility (MODS) culture with microscopy and solid culture for diagnosis of TB meningitis and HIV-associated pulmonary tuberculosis (TB). Study Design: Comparative study. Place and Duration of Study: Department of Clinical Pathology, Hasan Sadikin Hospital, Bandung, Indonesia, between 2010 and 2012. Methodology: Two groups of patients were included. The first group consisted of 167 consecutive HIV-infected patients presenting with suspected pulmonary TB. The second group consisted of 88 patients with clinical suspicion of TB meningitis. Sputum samples from HIV-associated pulmonary TB patients and cerebrospinal fluid (CSF) from patients with TB meningitis were analyzed using microscopy of Ziehl-Neelsen (ZN) stained smears, culture on solid medium (Ogawa), and MODS culture. Results: MODS showed the highest detection rate in both patient groups. Among HIV-associated pulmonary TB patients, positivity of MODS was 31.2% compared with 26.9% for Ogawa and 20.6% for ZN. Among TB meningitis patients, positivity of MODS was 41.2% compared with 38.8% for Ogawa and 8.3% for ZN. The median time to culture positivity was significantly shorter for MODS compared to Ogawa, both for sputum (median 11 vs 21 days) and CSF (14 vs 33 days). In 14 days, MODS detected significantly more cases compared with Ogawa in both patients group (79.2% vs 2.4% and 68.6% vs 0%, respectively). Laboratory staff readily used MODS after two weeks of training. Conclusion: We were able to implement MODS culture as a robust, sensitive, and rapid method for diagnosis of HIV-assoicated pulmonary TB and TB meningitis in a hospital setting in Indonesia. Further studies may be needed to assess the feasibility of MODS culture in other settings and assess its impact on case detection and timely treatment of both forms of TB.

Genetic variations and relationship among dengue virus type 3 strains isolated from patients with mild or severe form of dengue disease in Indonesia and Thailand.

Sequence analysis was conducted on structural and non-structural genes of 7 strains of dengue virus type-3 (DENV-3 virus) isolated in Indonesia and Thailand in the year 1973, 1994, and 1998 from patients with different clinical manifestations. In general, sequence similarity among isolates was greater than 93%, indicating that the mutation rate of DENV-3 circulating in this region was not more than 7% in the last 3 decades and suggesting that sequences that may responsible for viral architectures and/or biological function were strictly conserved. Mutations unique to viral strains associated with specific clinical manifestations were not found. Alignment of PrM/M and E nucleic acid sequences followed by parsimony analysis of sequences obtained in this study and published elsewhere allowed generation of phylogenetic trees, demonstrating that DENV-3 strains isolated in Indonesia in 1998 belonged to a separate cluster (subtype 2) from those isolated between 1973-1985 (subtype 1).

Implementation of 25-well culture plates for M. tuberculosis drug susceptibility testing in Indonesia.

At present, there is no standardized method for Mycobacterium tuberculosis drug susceptibility testing (DST) among laboratories in Indonesia. Since January 2001 to January 2004 we have tried to establish the method of 25-well culture plates with middlebrook’s media (Drug Susceptibility Culture Plate (DSCP) method) used by the Dutch Supranational Reference Laboratory at the Institute of Public Health and the Environment (RIVM), Bilthoven, Netherlands. Our experience showed that this method potentially gives better result as it can be very well standardized, faster and provides detailed MIC (Minimal Inhibitory Concentration) values. Data from 364 isolates that have been tested by DSCP method showed that resistance to INH, rifampicin, ethambutol, and streptomycin were 21.4%, 19.8%, 15.7%, and 16.5% respectively. Multidrug resistance were found in 13.2% isolates.

Enhancing knowledge and awareness of dengue during a prospective study of dengue fever.

In 1992, the Indonesian CDC implemented strategies to control and prevent dengue fever (DF) by including community involvement to reduce larva breeding sites and a mass health education program. To contribute to this effort, we incorporated an educational component into a prospective study of DF conducted at two textile factories in Bandung. This education provided: a lecture on the signs and symptoms of dengue and ways to prevent the disease, posters in the health clinic at each factory and handouts given to each volunteer with an explanation of symptoms. Upon enrollment, each participant completed a questionnaire to gather demographic information. Additionally they were given a brief (non-standardized) test (PRE-test) of their dengue knowledge, which was verbally administered by the study physicians. Five questions (15 point system) were designed to assess the participant's ability to recognize and describe aspects of dengue in lay terms. The subject material included: the symptoms of acute DF, transmission of dengue virus, and basic steps for disease prevention. The same questionnaire was re-administered 18 months later (POST-test), and the results were compared. A total of 2,340 participants completed both the PRE- and POST-tests; there were 1,373 males and 967 females, median age 36 years (range 18-59). Only 0.3% of participants scored EXCELLENT (15-14 points) on the PRE-test whereas 8.4% scored EXCELLENT on the POST-test. Fewer participants scored VERY BAD (2-0 points) on the POST-test compared to the PRE-test (1.4% vs 4.0%). The average raw scores for the PRE- and POST-tests were 7.8 and 10.1, respectively. Improvement of individual scores correlated highly with educational level. No significant correlation was identified for gender, age, factory location or a diagnosis of dengue during the study. These findings demonstrate that our prospective study enhanced knowledge and awareness of dengue in the volunteers.