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Hepatitis B surface antigen (HBsAg) is the earliest and most important serological marker for the diagnosis of HBV infection. The availability of new methods with a high sensitivity to detect HBsAg results in the increase of false reactive results so that a confirmatory test is needed,but this will increase the total test cost. A reactive cut-off value for a confirmatory test is needed to make the use of this test more efficient. This study was a cross-sectional. All the specimens with HBsAg >0.17 Cut-Off Index (COI) were confirmed with HBsAg confirmatory test. HBsAg test used a sandwich ELFA method while HBsAg confirmatory test used an antibody neutralization method. Analysis of the ROC curve obtained HBsAg cut-off value that need confirmatory test. Total samples were 80 with 51 (63.8%) confirmed reactive and 29 (36.2%) non-reactive. There was a statistically significant difference between HBsAg that confirmed reactive (median 2.76 COI) and non-reactive (median 0.32 COI) (p<0.001). ROC curve showed an AUC of 0.805 which meant a good diagnostic performance for HBsAg test based on a confirmatory test. The specificity of 89.66% and sensitivity 64.71% were obtained from the cut-off 1.08 COI and considered the best cut-off. Some possible causes of false reactive results were Hepatitis B vaccine, G-CSF therapy and limitation of the HBsAg methods. HBsAg cut-off with ELFA method that need HBsAg confirmatory test was <1.08 COI. The researchers suggests further studies with different sampling methods so a better data distribution can be obtained.

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Nephrotic syndrome (NS) is characterized by proteinuria, hypoalbuminemia accompanied by oedema and hypercholesterolemia.Nephrotic syndrome is an often relapsing disease (75%) and often the diagnosis is too late. This disease is 15 times greater in childrenthan in adult and the male to female ratio is 2:1. Laboratory examination is needed to rapidly detect and evaluate the progress of thedisease for treatment. To know the distribution of NS based on aged, gender, cholesterol, and albumin concentration and the urinesediment profile. The data in this retrospective descriptive study were collected from 56 patients with NS at the Wahidin SudirohusodoHospital, Makassar, in the period of January 2004 – June 2006. NS were found in 36 male patients (64.3%) and 20 female patients(35.7%). Cholesterol concentrations were 250 mg/dl in 50 patients (89.3%) and 250 mg/dl in 6 patients (10.7%). Albuminconcentration was 2.5 g/dl in 21 patients (37.5%) and < 2.5 gr/dl in 35 patients (62.5%). In urine sediments, there were found in 23patients (41.1%) with positive proteinuria (+++), 51 patients (91.1%) with positive erythrocytes, 54 patients (96.4%) with positiveleucocytes, and 33 patients (58.9%) with positive cylinders such as rugged granular and fatty cost. More NS were found in male patientsin comparison to female, and many were aged + 6 years. Hypercholesterolemia, hypoalbuminemia, proteinuria, hematuria, leucocyturia,and positive cylinder (rugged granular and fatty cast) were found in the urine of most of the NS patients.

BACKGROUND: Early diagnosis of tuberculosis (TB) cases in limited resource remains challenging. It is urgent to identify the new diagnostic tools which can control the spread of disease with accurate and rapid test. AIM: This study aimed to investigate the levels of infection markers: Composite bacterial infection index (CBII) and serum amyloid A (SAA) protein in pulmonary TB (PTB), and their healthy household contacts, as the alternative diagnostic markers for TB. METHODS: CBII and SAA were measured from 44 new PTB patients, and 31 household contact serum samples. The value of CBII was calculated from neutrophils, lymphocytes, monocytes, erythrocyte sedimentation rate, and high-sensitivity C-reactive protein (hs-CRP) level. hs-CRP and SAA levels were quantified from their serum samples using ELISA. QuantiFERON-TB Gold Plus (interferon gamma release assay [IGRA]) was used to screen latent TB infection among household contacts. RESULTS: Among 31 household contacts, there were 24 positive IGRA results and the rest (n = 7) had negative results. PTB patients exhibited significantly higher level CBII in the serum specimens, than those in household contact (p < 0.0001). There was no significant difference in the SAA level between TB cases and household contacts (p = 0.679). CONCLUSIONS: CBII can be used as one of the biomarkers for the identification of PTB from the serum specimens.

Pathogenic mycobacteria are one of the major causes of human mortality in the word. Mycobacterium tuberculosis is an etiological agent of human tuberculosis. Designing new vaccines including recombinant protein vaccines may be considered as a new approach for preventing or reducing tuberculosis epidemics. In order to construct protein recombinant as candidate vaccine, the Rv1980c gene encoding MPT64 protein was amplified from M. tuberculosis H37Rv strain genomic DNA using the PCR method and inserted into the cloning vector pGEM-T Easy. The recombinant plasmid pGEM-T Easy-MPT64 was then transformed into E. coli JM109 and cultivated under standard procedure, followed by plasmid extraction, PCR amplification, and DNA sequencing. The correct Rv1980c gene was confirmed by DNA sequencing and subcloned into expression vector pQE30Xa to yield recombinant plasmid pQE30Xa-MPT64, and transformed into E. coli BL21 strain. Transformed white recombinant colony was selected, cultured, induced with 40 μM IPTG, and identified using SDS-PAGE electrophoresis method. The molecular weight was found to be about 24 kDa and identified as recombinant protein MPT64. The target gene has been cloned into host E. coli BL-21 strain and expressed successfully as a soluble protein. The recombinant fusion recombinant protein MPT64 paves the way for tuberculosis diagnosis and vaccine development in the future, especially in Indonesia.

Increased resistance to TB drugs, may render vaccine development a more effective approach to stop or reduce TB epidemics. The antigen Culture Filtrat Protein Filtrat 21 (CPF21) is an immudominant protein encoded in RD 2 region of the Mycobacterium tuberculosis genome, capable of obtaining a strong hypersensitivity reaction and to induce very high interferon-gamma (IFN-γ) responses in patients with tuberculosis. In order to construct the recombinant plasmid pGEM-T Easy-CFP21 and express it in E. coli BL21, the CFP21 gene was amplified from M. tuberculosis H37Rv genomic DNA using PCR in vitro, and inserted into the pGEM-T Easy cloning vector. The recombinant plasmid was then transformed into E. coli JM109, followed by plasmid extraction, PCR amplification, and DNA sequencing. The correct recombinant CFP21 gene was subcloned into expression vector pGEX-2TK and transformed into E. coli BL21 strain. The white recombinant colony was selected, cultured, induced with 50 µM IPTG, and identified using SDS-PAGE electrophoresis method. These results demonstrated that CFP21 gene has been constructed and expressed successfully. The molecular weight was about 47 kDa as the fusion protein GST-CFP21 and expressed as insoluble protein. In conclusion, the target gene CFP21 has been cloned into host E. coli BL-21 strain and expressed successfully. In the future, the purified recombinant fusion protein GST-CFP21 paves the way for TB diagnosis and vaccine development.