Comparison between IS6110 and MPB64 primers for the diagnosis of Mycobacterium tuberculosis in Bangladesh by polymerase chain reaction (PCR).

The study was carried out to evaluate the efficacy of two different primers in PCR based diagnosis of tuberculosis on the basis of the time involvement, steps of PCR, cost and accuracy of results. Among 17 clinically suspected pulmonary tuberculosis patient's (M:F 65:35) sputum samples, 12 were found positive by both primers. Among 81 routine samples (sputum = 12; blood = 27; cerebrospinal fluid = 16; ascitic fluid = 12 and urine = 14), 12 were found to be positive by IS6110 primer while 13 were found positive by MPB64. The result of M. tuberculosis DNA PCR in sputum, ascitic fluid, CSF and urine samples were found to be similar in both the primers used. It is suggested that, M. tuberculosis DNA PCR in blood samples using MPB64 primers sometime gives false positive result and therefore, may not be recommended for blood samples. As in other samples both the primers gave almost the same result and therefore, both the primers can be used for MTB DNA PCR vice versa in order to cross check if one gives dubious result.

Urinary iodine status and thyroid dysfunction: a Bangladesh perspective.

Iodine deficiency is endemic in Bangladesh. Compulsory iodization of table salt was introduced since 1993 to prevent and improve thyroid disorders in the country. Urinary iodine status, thyroid function and antithyroid antibodies were studied in 397 newly diagnosed thyroid patients and 94 age-sex matched controls. Among thyroid patients, 96 were hyperthyroid, 185 euthyroid and 116 hypothyroid. Mean and median urinary iodine were higher (p=0.075) in thyroid patients (26.13+/-0.91 and 23.03) than controls (22.65+/-1.47 and 18.59); in hyperthyroid and euthyroid than hypothyroid (p=0.020); in multinodular (28.08+/-2.80 and 26.94) and diffuse (27.35+/-1.19 and 26.71) goitre than uninodular (23.91+/-2.37 and 19.14) and nongoitrous (NG, 21.5+/-2.05 and 18.27) (p=0.098) patients but no sex difference (p=0.466). Antimicrosomal (26.7%) and antithyroglobulin (34%) antibodies were more frequently positive among thyroid patients than controls (6.4% and 12.8% respectively) (p=0.00002 and p=0.00005 respectively). Antibody positivity was higher in diffuse (82/228) and multinodular (20/47) goitre than nongoitrous (20/56) and uninodular (13/66) goitre (p=0.046) as well as in hypothyroid (55.2%) and hyperthyroid (36.5%) than euthyroid (19.5%) patients (P<0.001). Urinary iodine correlated neither with antimicrosomal (thyroid patients: p=0.597 and control: p=0.112) nor with antithyroglobulin (thyroid patients: p=0.388 and control: p=0.195) antibody. Thyroid autoimmunity and dysfunction seems common; and interaction of salt iodization with iodine status and thyroid disorders may be important in Bangladesh.

Chronic carriers of hepatitis B virus in Bangladesh: a comparative analysis of HBV-DNA, HBeAg/anti-HBe, and liver function tests.

Serological markers of hepatitis B virus (HBV), liver function tests and quantitative estimation of HBV-DNA are important in the assessment of the state of infection and prognosis following treatment for hepatitis B. This study aimed to determine whether low-cost assays, eg hepatitis B e antigen (HBeAg) and liver function tests, could be used for the assessment of infectivity as an alternative to HBV-DNA estimation. We tested 125 hepatitis B carriers for HBeAg, antibody to HBeAg (anti-HBe), and serum HBV-DNA; we also carried out a range of standard liver function tests. Seventy-three subjects were positive and 52 were negative for HBeAg. Of the HBeAg positive cases, 3 were also positive for anti-HBe; of the HBeAg negative cases, 5 were also negative for anti-HBe. Of these 8 cases, 7 had no detectable HBV-DNA. Most of the HBeAg positive but anti-HBe negative subjects were positive for HBV-DNA (74.3%; 52/ 70) whereas most of the HBeAg negative and anti-HBe positive subjects (93.6%; 44/47) were also negative for HBV-DNA. Of 56 HBV-DNA positive individuals, alanine transaminase (ALT) was found to be raised in 69.6% (p=0.066) and aspartate transaminase (AST) was raised in 66.1% (p=0.011), while 67.9% had normal alkaline phosphatase (ALP) (p=0.054). HBeAg (p=0.018) and raised ALT (p=0.008) were found to be independent predictors for HBV-DNA positivity among HBV carriers. This study suggests that HBeAg positive and anti-HBe negative hepatitis B carriers with raised ALT and AST are likely to be positive for HBV-DNA; the combination of routine serology and biochemical tests may be considered as an alternative to HBV-DNA in evaluating the state of chronic HBV infection. However, HBV-DNA should be specifically assessed if discordance is observed between seromarkers and transaminases.

Predominance of the DEN-3 genotype during the recent dengue outbreak in Bangladesh.

A recent outbreak of dengue in Bangladesh was marked by many fatal complications. As clinical virulence varies among the genotypes of dengue virus, a study was conducted to investigate the molecular genotypes of dengue in Bangladesh. Reverse transcription polymerase chain reaction was used to determine viral genotypes using oligonucleotide generic primers that produce a 511 bp product. The resulting product was typed by nested PCR with strain-specific primers, yielding 482 (DEN-1), 119 (DEN-2), 290 (DEN-3) and 392 (DEN-4), visualized on UV transilluminator after electrophoresis on 2% agarose gel stained with ethidium bromide. Of 45 clinically diagnosed dengue patients (mean age 28 years; male/female 30/15), 19 (42.2%) had detectable viral RNA in their blood. However, during the first 5 days of fever in 30 patients, the frequency was 60% (18/30), implying that the sooner serum is drawn after the fever, the greater the chances of detecting viral RNA. DEN-3 was detected in all except 2 patients who were infected with DEN-2. DEN-2 (two cases) and DEN-4 (one case) were present as co-infections with DEN-3. All of the patients presented with fever, anorexia and vomiting; many had headache and general body ache; a few had a rash. About a quarter had suffered episodes of bleeding, while ascites, pleural effusion and CNS symptoms were found in a few patients Patients positive for viral RNA were also positive for anti-dengue IgM (p=0.007) in subsequent sampling. The study suggests the predominance of DEN-3 infection with occasional co-infection with other types, during the recent outbreak of dengue in Bangladesh.

Genetic background of diabetic and nondiabetic sibs in young Bangladeshis.

The prevalence of diabetes mellitus in the young is higher in Bangladesh like other Asian developing nations. Albeit, undernutrition has been shown to be associated with diabetes in the young, not all such individuals are diabetic. Diabetes Mellitus is a multigenic disease. In IDDM, DR3/4 heterozygotes were shown to have a greatly increased risk of developing the disease, suggesting the concept of genetic factor(s) being involved in the development of diabetes. Therefore, this study was undertaken to determine the distribution of HLA class II alleles (DRB) and to identify the HLA associated risk for developing diabetes mellitus in the young Bangladeshis. A total of fifty individuals were investigated. Half of them (n=25) were diabetic patients, registered in BIRDEM and half the participants were their non-diabetic sibs. A genomic DNA PCR and Enzyme Linked Probe Hybridization Assay (ELPHA, Bio-test, Germany) was used to determine HLA class II alleles (DRB1, DRB 3, 4, 5) by in vitro amplification of DRB gene. Among all the sero-equivalent antigens found in the study subjects, the prevalence of DR15 (DR2) was overrepresented, both in the diabetic subjects and in their non-diabetic sibs. Moreover, compared with the non-diabetic group the diabetic patients showed higher frequency of DR15 alleles (39 and 25%) though the difference was not significant (chisq. 1.7, p>0.05). Next to DR15, DR4 was the most prevalent HLA-DRB gene found in the study population. Interestingly, the frequency of DR4 was higher in the diabetic than in the non-diabetic group (20 vs. 14%). The study showed that the DR15 and DR4 were the most prevalent in the study population. Moreover, DR7 though not very significant, was higher in non-diabetic compared to their diabetic sibs. Comparison between the diabetic and non-diabetic sibs could have been interesting and significant but we could not confirm our findings, possibly, due to small sample size. A study in a larger paired sample of unrelated population is also needed to substantiate our findings, and also to prove the susceptibility or resistant haplotype in the young diabetic subjects.

Prevalence of infectious diseases and drug abuse among Bangladeshi workers.

Individuals seeking jobs abroad need health fitness certificates before entering into those countries. Medical screening of 43,213 Bangladeshi job seekers (M/F: 42,290/923) was carried out in our reference center during the period August, 1994 to May, 1996. Albeit male predominance, they represented middle and lower middle socio-economic class of the population from all over the country. All were young adults (age: 27.05+/-3.56 years; mean+/-SD) applying for job visas to different Asian countries. Physical examination and laboratory investigations including markers for several infectious diseases and drugs of abuse were carried out as required by countries recruiting the workers. Serological tests revealed that 1,884 (4.4%) of individuals were positive for hepatitis B surface antigen (HBsAg), 737 (1.7%) for Treponema pallidum hemagglutination (TPHA) and only 83 (0.2%) for antibody to human immunodeficiency virus (anti-HIV). However, we could not confirm any case of infection with HIV. Chest X-ray suggestive of pulmonary tuberculosis was found in 162 (0.4%) and on blood film, malarial parasites could be observed only in 4 cases. Their urine analysis revealed the presence of opiates or cannabinoids in 471 (1.1%) individuals. HBsAg-positive cases (p = 0.003) and abuse of opiates (p = 0.024) or cannabinoids (p = 0.002) were significantly higher among males. TPHA reactivity and chest X-ray suggestive of tuberculosis were found to be higher among opiates (p = 0.002 and 0.027) and cannabinoids (p = 0.000 for both) abused as well as with increasing age (p = 0.000). These results may represent a cross-sectional view of the prevalence of different infectious diseases and abuse of drugs among the young adult population of Bangladesh.