Diarrheal diseases are the second cause of the high morbidity and mortality in children under five years old. According to the Basic Health Survey 2018 conducted by the Ministry of Health, the prevalence of diarrheal diseases among children under five years old that were diagnosed by healthcare workers was 11.0%. The aim of this study was to describe the enteric pathogen isolated from children with diarrhea. The study was conducted in five cities in Indonesia: Jakarta, Serang, Denpasar, Makassar, and Mataram. The Inclusion criteria were children aged one month to five years old, with diarrhea that was diagnosed by a healthcare worker. The rectal swabs were sent to the Centre for Research and Development for Biomedical and Basic Health Technology, National Institute of Health Research and Development, Ministry of Health in Jakarta. Virus and Enterotoxigenic Escherichia coli (ETEC) identification by using multiplex PCR from Seegene, meanwhile bacteria identified by conventional method. As many as 2626 children under five years old participated in this study. The highest viral pathogen that causes diarrhea is viral 1.807 (68,81%) and 486 (18,56%). The virus etiology was Rotavirus 982 (54,34%) cases, followed by Adenovirus 916 (50.69) cases, Norovirus II 444 (24,57%) cases, meanwhile the bacteria pathogen were Enterotoxigenic Escherichia coli detected in 262 (9,98%) followed by Campylobacter jejuni and Shigella spp. This study described Rotavirus is the prevalence etiology of diarrhea among children under five years old followed by Adenovirus and Norovirus, some other cases reported the cause of diarrhea were bacteria ETEC E. coli followed Campylobacter jejuni, Shigella spp, etc.
Pertussis cases have been reported most frequently in developed countries, but they are predicted to be the most prevalent in developing countries. Indonesia, a developing country, routinely conducts case-based surveillance for pertussis. We reviewed the data on pertussis cases and close contacts based on clinical sample documents examined in the National Reference Laboratory for pertussis, Indonesia (2016–2020). Our objective was to analyze the laboratory and epidemiological aspects of pertussis cases and close contacts, particularly to evaluate the implementation of a 5-year case-based surveillance of pertussis in Indonesia. Data were collected from sample documents and annual laboratory reports between January 2016 and December 2020. We analyzed the proportion of pertussis cases and close contacts by geographic region, year, age, and sex. We used the χ2 test to correlate the laboratory and epidemiological data. In total, 274 clinical cases of pertussis and 491 close contacts were recorded in 15 provinces. The peak number of cases occurred in 2019, with a positivity rate (percentage of laboratory-confirmed cases) of 41.23% (47/114). Clinical cases were dominated by infants aged <1 year (55.5%), and 52.9% of them were aged <6 months. Similarly, 72.3% (68/94) of the laboratory-confirmed cases were infants. Both clinical cases and positivity rates tended to be higher in females (155 cases, 38.1%) than in males (119 cases, 29.4%). No confirmed cases were found in children aged ≥10 years, although positive results still occurred in close contact. Age-group and laboratory-confirmed cases were correlated (p = 0.00). Clinical and confirmed cases of pertussis occurred mostly in the early age group and may be lower in those aged ≥10 years, especially in confirmed cases. New policies are needed for pertussis prevention at an early age, as well as the application of serology tests to increase laboratory-confirmed cases in children aged ≥10 years.
Indonesia is one of the five countries with highest diphtheria cases in the world. Laboratory confirmation by culture method as a gold standard requires bacterial survival. Indonesia’s geographical condition as an archipelagic country and difficulties in transporting clinical samples are often obstacles in maintaining bacterial survival. This study aims to evaluate the ability of several transport mediums to maintain the survival of Corynebacterium diphtheriae. A total of 90 isolates were divided into nine groups of transport mediums. Samples were divided into 2 treatment groups, namely room temperature and temperature 2-8 ºC. On day 2, 4, 8, 16, and 32, 1 isolate from each group with 2 different incubation temperatures was cultured on blood agar medium and incubated for 24 hours at 37 ºC. Bacterial survival was indicated by the growth of suspect colonies which were identified by microscopic and biochemical tests. Results show serum with tellurite can be used with viability lower than silica gel, but higher than other media. Meanwhile at a temperature of 2-8 ºC, there are 2 types of the best transport medium, namely serum with tellurite and open silica gel in aluminum foil. Newborn Calf Serum supplemented with Tellurite can be used as an alternative transport medium for Corynebacterium diphtheriae, both at room temperature and at 2-8 ºC.
Diphtheria is a vaccine-preventable disease. The clinical features and complications of diphtheria are associated with toxins produced by the causative bacteria. Diphtheria toxin synthesis is encoded by tox gene. This study aimed to provide an overview of the DNA sequences of the tox gene of Corynebacterium diphtheriae causing diphtheria in several region of Indonesia. A total of 65 Corynebacterium diphtheriae isolated from several provinces in Indonesia (2010-2017) were used as samples. Isolates recultured on blood agar medium (BA), incubated at 37 0 C overnight. DNA extraction conducted using the QiaAmp DNA Mini Kit. The DNA sequencing was carried out using the Whole Genome Sequencing (WGS) approach. The data conversion and analysis conducted using U-gene and BioEdit programs. Examination of 65 isolate C. diphtheriae with 1683 bp of tox gene sequences showed that there are 3 patterns of gene sequences with 3 mutation site. All mutations were silent mutation. The mutation sites were also not commonly used as 3’end binding site of the PCR primer. We concluded that tox gene of C. diphtheriae that causes diphtheria in some provinces in Indonesia have limited variations and these variations do not encode amino acid changes. This indicates that the vaccines used in Indonesia are still in accordance with the variations in circulating bacteria and PCR can be used for screening and predicting the toxigenicity of diphtheria-causing bacteria. Keywords: C. diphtheriae, gene tox, diphtheria, Indonesia Abstrak Difteri merupakan salah satu penyakit yang dapat dicegah dengan imunisasi (PD3I). Gambaran klinis dan komplikasi difteri dikaitkan dengan toksin yang diproduksi oleh bakteri penyebab. Sintesis toksin difteri dikode oleh gen tox. Penelitian ini bertujuan untuk memberikan gambaran sekuens DNA gen tox Corynebacterium diphtheriae penyebab difteri di beberapa wilayah Indonesia. Sebanyak 65 isolat C. diphtheriae tersimpan milik Badan Litbangkes yang diisolasi dari beberapa wilayah Indonesia tahun 2010- 2017 dijadikan sebagai sampel. Rekultur dilakukan pada medium agar darah (BA), diinkubasi pada suhu 37 o C selama sehari semalam. Ekstraksi DNA menggunakan kit QiaAmp DNA Minikit. Sekuensing DNA dilakukan dengan pendekatan Whole Genome Sequencing (WGS). Konversi dan analisis data menggunakan program U-gene dan BioEdit.Pemeriksaan 65 isolat C. diphtheriae dengan 1683 bp sekuens gen tox menunjukkan ada 3 pola sekuens gen dengan 3 lokasi mutasi. Seluruh mutasi bersifat silent mutation. Lokasi mutasi juga bukan merupakan tempat penempelan ujung 3’ primer PCR yang umum digunakan. Berdasarkan hasil penelitian dapat disimpulkan bahwa variasi gen tox yang ditemukan pada C. diphtheriae penyebab difteri di Indonesia memiliki variasi yang terbatas dan mutasi yang ada tidak mengkode perubahan asam amino. Hal ini mengindikasikan bahwa vaksin yang digunakan di Indonesia masih sesuai dengan variasi bakteri yang bersirkulasi. Hasil penelitian juga mengindikasikan bahwa PCR dapat digunakan untuk skrining dan memprediksi toksigenisitas bakteri penyebab difteri. Kata kunci : C. diphtheriae, gen tox, difteri, Indonesia
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