The First Case of Laboratory-confirmed Dengue Virus Infection in Mimika, Papua Province, Indonesia

Background: Dengue is the most important vector-borne disease that poses serious health problem both in tropical and subtropical countries. Since the first outbreak in Surabaya in 1968, dengue infection has spread in all provinces in Indonesia. Mimika district in Papua province, Indonesia, is a non-endemic dengue area with no laboratory-confirmed case reported. However, until 2012 there were 13 suspected dengue infection admitted to local General Hospital in Mimika district, Papua province, Indonesia. This study described the genetic characteristics of first laboratory-confirmed dengue virus (DENV) infection in Mimika district, Papua province, Indonesia. Methods: Viral isolation in C6/36 cell line, RT-PCR and serotyping were carried out to confirm the presence of DENV within serum patient of suspected DENV cases from Mimika district, Papua Province, Indonesia. Direct sequencing and phylogenetic analysis of complete coding sequence (CDS) of E gene was performed to the samples that have already confirmed positive DENV for viral genotyping. Results: Four cases were confirmed to be DENV by RT-PCR while only 2 samples were able to be culture in C6/36 mosquito cell line. Serotyping confirmed that the DENV from Mimika district, Papua province were DENV3 serotype. The genotyping showed that the DENV3 from Mimika district were belonged to genotype I. Conclusion: This study reported the first laboratory-confirmed of DENV cases in non-endemic dengue area, Mimika district, Papua Province, Indonesia.

DENV has caused health public problem since its ¿UVW RXWEUHDN UHSRUW LQ 6XUDED\D LQ ZLWK D WRWDO RI FOLQLFDO FDVHV DQG GHDWKV In addition, the largest incidence of DF and DHF in the South East Asia region since 2004 was reported in Jakarta. 10 Nowadays, DENV infection has occurred in all provinces in Indonesia. 11 Mimika district in Papua has been recognized as non-endemic area of DENV since no laboratory-FRQ¿UPHG FDVHV ZHUH UHSRUWHG +RZHYHU LQ mid-2012, there were suspected cases of dengue reported to local Hospital and the specimens were sent to National Institute of Health Research and 'HYHORSPHQW RI +HDOWK 1,+5' -DNDUWD IRU GHQJXH virus examination and genetic characterization. The genetic information of these viruses is essential since these viruses obtained from non-endemic area.
This report described the genetic characteristics of WKH ¿UVW FDVHV RI GHQJXH IRXQG LQ 0LPLND GLVWULFW Papua Province. Molecular analysis of dengue virus using bioinformatic analysis was performed for genotyping of the DENV from Mimika district, Papua Province.

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In June 2012, Virology laboratory, NIHRD, received 13 serum samples of suspected dengue cases from +HDOWK RI¿FH RI 0LPLND 'LVWULFW 3DSXD 3URYLQFH $OO of the serum samples were obtained from suspected dengue patients admitted in local General Hospital in Mimika District. Four out of 13 serum samples were positive dengue IgM and IgG by rapid diagnostic test performed at the local General Hospital in Mimika District. However, NIHRD only received the specimens ZLWKRXW DQ\ LQIRUPDWLRQ IURP WKH +HDOWK RI¿FH GLVWULFW The demographic data such as age, gender, address and date of onset were not available from all of the samples. This become the limitation of this report as the epidemiological analysis could not be performed. All of the serum samples received in NIHRD, Jakarta, were lack of complete demographic data such as date of onset or detailed clinical symptoms'. Therefore demographic data was not described in this report.

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Dengue infection has spread in all provinces in Indonesia and caused outbreaks annually. Several studies had reported dengue outbreak in several cities in Indonesia in which both young age and older age groups were infected by DENV and KDG GHQJXH KHPRUUKDJLF IHYHU '+) LQFLGHQFH 13 Mimika district is one of districts from Papua province, located in Irian Island, the Eastern part of ,QGRQHVLDQ DUFKLSHODJR )LJ 7KLV DUHD LV NQRZQ as non-endemic area of dengue since there was no GDWD RI ODERUDWRU\ FRQ¿UPHG GHQJXH FDVHV UHSRUWHG from Mimika district, Papua province. Here we UHSRUWHG WKH ¿UVW FDVHV RI ODERUDWRU\ FRQ¿UPHG DQG genotyping of DENV taken from dengue outbreak in Mimika district, Papua Province, Indonesia, in 2012.

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Suspected samples from Mimika district were FRQ¿UPHG WR EH '(19 JHQRW\SH , LQ ZKLFK WKLV serotype is predominant in other cities in Indonesia along with DENV-2. 11 The DENV-3 genotype I was already found in other cities in Indonesia 14,15 and also commonly found in South East Asia. 7 The Srt_Mimika2012 was clustered closely with other sequences from Indonesia, therefore it was possible that the Mimika virus was imported from other place VLQFH RU EHIRUH 3K\ORJHQHWLF DQDO\VLV FRQ¿UPHG WKDW WKH JHQRW\SH distributions of DENV strains circulating in each of the South East Asian Countries remained stable. DENV-3 genotype I was mainly distributed in South (DVW $VLD DQG 3DFL¿F DOWKRXJK VRPH VWXGLHV DOVR reported genotype III virus was introduced into South East Asian Countries and Southern China. This virus had caused dengue outbreak in recent years. 16 7KH ODERUDWRU\ FRQ¿UPDWLRQ DQG JHQRW\SLQJ RI DENV from Mimika, Papua province, has informed KHDOWK RI¿FHU DQG JRYHUQPHQW DXWKRULWLHV FRQFHUQLQJ the DENV circulation in Mimika, Papua Province. According to the data obtained from Center Disease Control and Prevention, Ministry of Health, Republic of Indonesia, in 2012 to 2014 the incidence rate of dengue infection in Papua province is low. This data ZDV SURYLGHG ZLWK GLVWULFW KHDOWK RI¿FHUV EDVHG RQ dengue cases found in primary health care or local hospital. However, since Malaria and HIV is the main health problem in Papua, dengue cases were not considered as priority in Papua province. Therefore the reported cases were low and many dengue infections were undetermined or under-reported.

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hospital is only based on clinical symptoms and trombocytopenia in which could be triggered by other viruses and may leads to missed diagnose. 7KHUHIRUH LW LV QHFHVVDU\ WR FRQ¿UPHG WKH GHQJXH suspected cases using laboratory testing for DENV DQWLJHQ RU '(19 VSHFL¿F DQWLERG\ GHWHFWLRQ Among four samples out of 13 samples that were positive for DENV, only two samples were able to grow in C6/36 cell line and only one complete coding sequence of E gene was obtained from one sample. The possible reason for this is the low virus concentration in sera samples due to time of samples collection. The samples collection exceeding the acute phase where viremia had already passed can cause low titer of virus and affecting the examination results. However the disease phase from each patient in this report could not be determined since the date of onset was not available.
The authors acknowledged the lack of epidemiology data from each patient became the major limitation in this study. It is not known whether the cases were imported or emerge locally since address or travel history of patient admitted to the local General Hospital in Mimika District were not available. The epidemiological data such as the date of onset and GHPRJUD¿F GDWD PD\ SURYLGH EHWWHU XQGHUVWDQGLQJ RI the distribution of DENV in Mimika district, Papua province.

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We acknowledge the Government of Republic of Indonesia for the funding. We also thank district KHDOWK RI¿FHU LQ 0LPLND 3DSXD SURYLQFH IRU providing the data in this study.