Yati Soenarto
Departments of Pediatric, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta Department of Pediatric, Dr.Sardjito General Hospital, Yogyakarta

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Genotyping of Rotavirus by Using RT-PCR Methods Nirwati, Hera; Wibawa, Tri; Aman, Abu Tholib; Soenarto, Yati
Indonesian Journal of Biotechnology Vol 18, No 1 (2013)
Publisher : Universitas Gadjah Mada

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Abstract

There is a great diversity of rotavirus genotypes circulating worldwide, with dominant genotypes changing from year to year. Rotavirus genotyping was performed by using reverse transcription PCR with type-specifi c-primers. Since rotavirus is a RNA virus that has high mutation rate, there was a possibility of technical diffi culty in genotyping due to mutation in the primer binding sites. During Indonesian rotavirus surveillance study 2006-2009, it was reported that 17% of samples subjected for G type and 21% of samplessubjected for P type were untypeable. The objective of this study was to identify genotypes of the samples that were untypeable previously using RT-PCR based on the method described by Das et al. (1994) and Gentsch et al. (1992). There were 30 samples subjected to G type and 61 samples subjected to P type to be re-typed using method described by Gouvea et al. (1990) and Simmond et al. (2008) for G and P typing, respectively. By using another set of primer, the genotype of all samples was identifi ed. This study highlights the importance of a constant reconsideration of primer sequences employed for the molecular typing of rotaviruses.Key words: rotavirus, G typing, P typing
Surveillance of Rotavirus Diarrhea in Dr. Hasan Sadikin General Hospital Bandung Prasetyo, Dwi; Martiza, Iesje; Soenarto, Yati
Majalah Kedokteran Bandung Vol 42, No 4
Publisher : Faculty of Medicine, Universitas Padjadjaran

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Abstract

The diarrhea morbidity in Indonesia has increased, however, all the reports had not been done carefully, so that accurate surveillance are essential for improving quality of morbidity data. To determine the prevalence and clinical manifestations of rotavirus diarrhea and to characterize the circulating rotavirus strains, children below 5 years old who were admitted to Hasan Sadikin Hospital, Bandung because of diarrhea, from January 2006 through March 2007 were enrolled in a surveillance study and had stool specimens tested for the presence of rotavirus using enzyme immunoassay (EIA). The strains of rotavirus were determined using reverse transcriptase-polymerase chain reaction (RT-PCR). Rotavirus were detected in 47.8% analyzed samples (87/184), G and P-genotype of rotavirus were G[1] (37.5%) and P[6] (53.5%). Most subjects were males (56%), 6–11 months of age (35%). Most common clinical manifestations besides diarrhea were dehydration (72.7%) and vomiting (50%). Subjects with positive rotavirus more common had dehydration (72% vs 28%) and vomiting (61% vs 39%). In conclusion, vomiting and dehydration are the prominent clinical manifestations of diarrhea with positive rotavirus infection. G1 and P6 are the most common genotype of rotavirus. [MKB. 2010;42(4):155–60].Key words: Clinical manifestations, diarrhea, genotype, prevalence, rotavirus Surveilans Diare Rotavirus di Rumah Sakit Dr. Hasan Sadikin BandungMorbiditas diare di Indonesia meningkat, tetapi semua laporan belum dilakukan secara cermat, sehingga surveilans yang akurat penting untuk memperbaiki kualitas data. Penelitian ini dilakukan untuk menentukan prevalensi dan manifestasi klinis diare rotavirus serta karakteristik strain rotavirus, anak usia di bawah 5 tahun yang dirawat karena diare di RS Hasan Sadikin, Bandung, dari Januari 2006 sampai Maret 2007, diikutsertakan dalam suatu penelitian surveilans dan spesimen fesesnya diperiksa untuk mendeteksi adanya rotavirus dengan menggunakan enzyme immunoassay (EIA). Strain rotavirus diperiksa dengan reverse transcriptase-polymerase chain reaction (RT-PCR). Rotavirus terdeteksi pada 47,8% sampel analisis (87/184), genotipe-G dan P adalah G[1] (37,5%) dan P[6] (53,5%) dari strain. Kebanyakan subjek adalah laki-laki (56%) usia 6–11 bulan (35%). Manifestasi klinis terbanyak adalah dehidrasi (72,7%) dan muntah (50%). Subjek dengan rotavirus positif lebih sering mengalami dehidrasi (72% vs 28%) dan muntah (61% vs 39%). Simpulan, muntah dan dehidrasi adalah manifestasi klinis terbanyak pada diare dengan infeksi rotavirus. Genotipe G1 dan P6 merupakan genotipe rotavirus yang paling sering ditemukan. [MKB.2010;42(4):155–60].Kata kunci: Diare, genotipe, manifestasi klinis, prevalensi, rotavirus DOI: http://dx.doi.org/10.15395/mkb.v42n4.29