Detection of Opportunistic Fungus Pneumocystis jirovecii Major Surface Glycoprotein (MSG) gene in HIV-AIDS Patients with Pneumoniae in Jakarta

Pneumocystis jirovecii is known to cause opportunistic infections in the lower respiratory tract in individuals with low immune systems, especially patient with HIV infection. The prevalence of P. jirovecii pneumonia (PjP) in various countries show varying numbers. In Indonesia, HIV cases continue to rise. However, the data in Indonesia concerning the case of PjP is very limited. Until now the prevalence of PjP in Indonesia is only based on clinical symptoms of the patient. Currently, diagnosis of PjP relies on microscopic examination. The disadvantage of this examination is not easy to do and has a high negative predictive value. Thus, this study was conducted to develop a molecular test to diagnose PjP infection in HIV-AIDS suspected pneumonia. Molecular diagnostic test aimed for Major Surface Glycoprotein (MSG) gene of P. jirovecii detection was done through real-time PCR against 100 sputum samples. Demographic data show that the prevalence of PjP infection in HIV-AIDS suspected pneumonia patients in Jakarta is 20.0%, male 75% within 31-40 y.o (35%), dominant (80%) from patients with CD4+ T-lymphocytes of 200-349 cells/µL. Molecular real-time PCR methods were shown to give five times sensitivity higher than Giemsa stain.   Keywords: P. jirovecii, HIV, real-time PC

The burden of diarrhoea, shigellosis, and cholera in North Jakarta, Indonesia: findings from 24 months surveillance

BACKGROUND: In preparation of vaccines trials to estimate protection against shigellosis and cholera we conducted a two-year community-based surveillance study in an impoverished area of North Jakarta which provided updated information on the disease burden in the area. METHODS: We conducted a two-year community-based surveillance study from August 2001 to July 2003 in an impoverished area of North Jakarta to assess the burden of diarrhoea, shigellosis, and cholera. At participating health care providers, a case report form was completed and stool sample collected from cases presenting with diarrhoea. RESULTS: Infants had the highest incidences of diarrhoea (759/1 000/year) and cholera (4/1 000/year). Diarrhea incidence was significantly higher in boys under 5 years (387/1 000/year) than girls under 5 years (309/1 000/year; p < 0.001). Children aged 1 to 2 years had the highest incidence of shigellosis (32/1 000/year). Shigella flexneri was the most common Shigella species isolated and 73% to 95% of these isolates were resistant to ampicillin, trimethoprim-sulfamethoxazole, chloramphenicol and tetracycline but remain susceptible to nalidixic acid, ciprofloxacin, and ceftriaxone. We found an overall incidence of cholera of 0.5/1 000/year. Cholera was most common in children, with the highest incidence at 4/1 000/year in those less than 1 year of age. Of the 154 V. cholerae O1 isolates, 89 (58%) were of the El Tor Ogawa serotype and 65 (42%) were El Tor Inaba. Thirty-four percent of patients with cholera were intravenously rehydrated and 22% required hospitalization. V. parahaemolyticus infections were detected sporadically but increased from July 2002 onwards. CONCLUSION: Diarrhoea causes a heavy public health burden in Jakarta particularly in young children. The impact of shigellosis is exacerbated by the threat of antimicrobial resistance, whereas that of cholera is aggravated by its severe manifestations

Correction: The burden of diarrhoea, shigellosis, and cholera in North Jakarta, Indonesia: findings from 24 months surveillance

This is a correction of an earlier published article

Diarrhoea Episodes and Treatment-seeking Behaviour in a Slum Area of North Jakarta, Indonesia

Visits to household during a census in an impoverished area of north Jakarta were used for exploring the four-week prevalence of diarrhoea, factors associated with episodes of diarrhoea, and the patterns of healthcare use. For 160,261 urban slum-dwellers, information was collected on the socioeconomic status of the household and on diarrhoea episodes of individual household residents in the preceding four weeks. In households with a reported case of diarrhoea, the household head was asked which form of healthcare was used first. In total, 8,074 individuals (5%)\uaf13% of children aged less than five years and 4% of adults\uafhad a diarrhoea episode in the preceding four weeks. The two strongest factors associated with a history of diarrhoea were a diarrhoea episode in another household member in the four weeks preceding the interview (adjusted odds ratio [OR] 11.1; 95% confidence interval [CI] 10.4- 11.8) and age less than five years (adjusted OR 3.4; 95% CI 3.2-3.5). Of the 8,074 diarrhoea cases, 1,969 (25%) treated themselves, 1,822 (23%) visited a public-health centre (PHC), 1,462 (18%) visited a private practitioner or a private clinic, 1,318 (16%) presented at a hospital, 753 (9%) bought drugs from a drug vendor, and 750 (9%) used other healthcare providers, such as belian (traditional healers). Children with diarrhoea were most often brought to a PHC, a private clinic, or a hospital for treatment. Compared to children, adults with diarrhoea were more likely to treat themselves. Individuals from households in the lowest-income group were significantly more likely to attend a PHC for treatment of diarrhoea compared to individuals from households in the middle- and higher-income groups

Differences of antigenic profiles on immunoblotting of wild type measles virus and vaccine virus in Indonesia

Background Measles virus has a negative, single strand RNA genome which codes for six important structural proteins. The genes of the wild type measles virus have many variances hence the nucleotide sequences of each wild type virus and vaccine virus are different. This differences lead to the antigenic differences between wild type and vaccine virus. Objective The purpose of this research is to investigate the differences in the antigenic profiles on immunoblotting between wild type and vaccine virus. Results The analysis results are 1) the antigen ofCAM-70 vaccine virus was less able in cross reacting with the antibodies from G2, G3, 09, CAM-70 and Schwarz; 2) The antibody aga inst CAM- 70 was only able to cross react with antigens of N protein and a few of antigens ofF proteins; 3) The wild type virus were very immunogenic, hence the antibody titers were very high; 4) The CAM-70 and MMR vaccine virus were less immunogenic, hence their antibody were very low; 5) The antibody responses that always occurred from all immunized mice serum were antibody for N and F proteins. However, the antibody against CAM-70 vaccine virus was still able to react with wild type virus (G2, G3 and 09). Conclusion All antigen-antibody reaction on immunoblotting resulted in different profiles especially between wild type virus and CAM-70 vaccine virus. Although CAM-70 vaccine virus showed clear differences compared to G2, G3 and 09 genotypes, antibodies against CAM-70 were still able to cross react with antigens from other genotypes (G2, G3 and D9)

Perbedaan Sekuens Asam Amino Epitop Sel B dan Sel T pada Protein Hemaglutinin (H) Antara Virus Campak Liar dan Virus Vaksin di Indonesia

Latar belakang. Protein H virus campak sangat penting agar virus dapat menginfeksi sel pejamu. Selain itu, protein H dapat merangsang antibodi spesifik yang dapat menetralisasi virus campak, sehingga virus tidak dapat menginfeksi sel. Bila ada perbedaan sekuens asam amino epitop sel B dan sel T pada protein H antara virus campak liar dan virus vaksin campak, maka vaksin tidak dapat merangsang terbentuknya antibodi protektif. Tujuan. Mengetahui perbedaan sekuens asam amino epitop sel B dan sel T pada protein H antara virus campak liar (G2, G3, dan D9) dan virus vaksin CAM-70, Schwarz dan Edmonston-wt. Metode. Ekstraksi dan amplifikasi gen dilakukan di laboratorium menggunakan teknologi biologi molekuler dan analisis gen dan protein dilakukan menggunakan teknologi bioinformatika. Hasil. Ditemukan perbedaan sekuens asam amino epitop sel T pada protein H antara virus campak liar dan virus Edmonstone-wt, sedangkan antara virus campak liar dan virus vaksin (CAM-70 dan vaksin Schwarz) tidak ditemukan perbedaan. Ditemukan perbedaan sekuens asam amino pada epitop sel B protein H antara virus campak liar dan virus vaksin (CAM-70 dan Schwarz, sedangkan antara CAM-70 dan Schwarz tidak ditemukan perbedaan. Kesimpulan. Tidak ada perbedaan sekuens asam amino epitop sel T antara virus campak liar dan virus vaksin (Schwarz dan CAM-70). Perbedaan ditemukan pada epitop sel B antara virus campak liar dan virus vaksin (CAM-70 dan Schwarz)

Critical site differences of fusion protein between wildtype and vaccine measles virus strains in Indonesia

Background Measles virus can cause high morbidity and mortality in infants and children. Fusion glycoprotein (F protein) found in the viral envelope is important for the host cell infection mechanism. F protein is immunogenic and may cause specific immune responses in the host. High variability is found in the F protein gene of vaccine viral strains compared to 'Wild type strains. This amino add sequence variability may result in a less specific immune response against other strains, possibly rendering the vaccine to be less effective. Objective To detennine the amino add sequence differences in critical sites of F protein in Mld type and vaccine measles virus strains in Indonesia. Methods We compared amino acid sequences of three genotypes of Mld type measles virus (02, 03 and D9) to those of the vaccine strains, CAM 70, Schwarz, and Edmonstonwt type measles virus. Resul ts Analysis showed that there were differences at FlF2 cleavage site, B cell epitopes, and H protein binding site between the CAM70 vaccine viral strains and Mld type strains. Schwarz vaccine strain differed from the wild type strains at the H protein binding site. A 03 wild type strain potential glycosylation site was also different from all other strains studied. Conclusion There were differences in the critical sites of F protein between Mld type strains and the CAM70 and Schwarz vaccine strains

Difference of hemaglutinins between wild-type and vaccine measles virus in Indonesia

Background Hemaglutinin (H) protein of measles virus is very important in the process of host cell infection. H protein is also able to induce specific antibodies which can neutralize measles virus and block the cell infection. Objective This study aimed to explore the nucleotide and amino acid sequence differences between wild-type measles virus (G2, G3 and D9) with CAM-70, Schwarz and Edmonston-wt vaccine virus. Methods The exctration and amplification of the gene were conducted in the laboratory using biomolecular technology. The gene and protein analysis were conducted using the bioinformatic technology. Results The results showed that the differences in nucleotide sequences were highest between wild-type virus and CAM-70 vaccine virus (76-77 nucleotides), followed by Schwarz (61-64 nucleotides) and Edmonston (60-63 nucleotides). The differences in amino acid sequences were highest between wild-type virus and CAM-70 (24-29 residues), followed by Schwarz (13-20 residues) and Edmonston (12-19 residues). Conclusion The Indonesian wild-type measles virus was genetically closer to Schwarz vaccine virus than CAM-70 vaccine virus, hence the neutralizing antibodies generated by Schwarz vaccine were more specific against Indonesian wild-type virus compared to CAM-70 vaccine

Antigenic differences between wildtype measles viruses and vaccine viruses in Indonesia

Background Measles virus has a single, negative strand RNA genome which codes 6 structural proteins: N, F, P M, H and L. Currently there are several variances in the nucleotide sequences of N, F, M and H genes across wild type measles viruses, hence measles viruses can be categorized into clades and genotypes. The antigenicity of the previous genotype of measles is different from the current genotype. Objective To determine the antigenic differences between wild type measles virus and measles vaccine virus. Methods Analysis of the antigenic differences between wild type virus (G2, G3 and D9) and vaccine virus (CAM-70 and Schwarz) was performed by immunizing mice with the respective viruses. The serum was then tested with micro-cross-neutralization technique using the G2, G3, D9 and CAM-70 virus. Tests with cross ELISA examination technique were also performed using the same set of virus. Results Analysis of the cross neutralization test and cross ELISA showed that the highest antigenicity reaction was found between wild type virus with antibody against wild type virus, while the lowest reaction was between wild type virus with antibody against CAM-70. Conclusions We conclude that the antigenicity of antigenic protein from wild type virus is higher than antigenicity of vaccine virus protein. In addition, it was found that the antigenicity of proteins from Schwarz vaccine virus was higher than proteins CAM-70 vaccine virus

Differences of nucleotide and amino acid sequences of nucleoprotein (N) gene between wild-type measles virus and vaccine virus in Indonesia

Background Measles virus is a member of genus morbilivirus which belongs to family paramyxovirus with negative, single- strand RNA genome. RNA is packed by nucleocapsid (N) protein. The N protein is very important for RNA replication and translation. Abnormality in N protein will induce abnormality in virus replication. Objective This study aimed to explore the differences of nucleotide sequence of N gene and amino acid sequences of N protein between wild-type measles virus (G2, G3 and D9) and vaccine virus (CAM-70, Schwarz and Edmonston-wt) Methods The exctraction and amplification of the gene were conducted in the laboratory using biomolecular technology. The gene and protein analysis were conducted using the bioinformatic technology. Results The results showed that more differences were found between nucleotide sequences of N gene of wild-type measles virus against CAM-70 vaccine virus (77 – 79 nucleotides) compared against Schwarz and Edmonston-wt vaccine virus (71- 74 nucleotides). Likewise, more differences were also observed between amino acid sequences of N protein of wild-type measles virus against CAM-70 vaccine virus (18-24 residues) compared against Schwarz and Edmonston-wt vaccine virus (17-23 residues)