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Abstrak :
Latar Belakang : COVID- 19 disebabkan SARS-COV-2. WHO menerbitkan protokol pemeriksaan laboratorium untuk deteksi virus menggunakan metode real time RT-PCR dari spesimen swab nasofaring dan orofaring. Metode ini cukup invasif. Diperlukan tehnik pemeriksaan yang relatif aman dan nyaman untuk pasien. Penelitian ini bertujuan untuk melihat efetivitas swab bukal sebagai alternatif pemeriksaan SARS-COV-2. Metode : Studi uji diagnostik ini dilaksanakan sejak tahun 2020 - 2021, mengambil spesimen swab nasofaring, swab orofaring dan swab bukal dari pasien positif COVID- 19. Dilakukan optimasi, ekstraksi RNA virus dan real time RT-PCR . Hasil Penelitian : Hasil studi mengumpulkan 68 spesimen dari pasien COVID-19. Hasil uji nasofaring, orofaring dan bukal positif adalah 24 spesimen. Hasil uji nasofaring dan orofaring positif dengan uji bukal negatif adalah 23 spesimen. Berdasarkan nilai Ct < 20 dan Ct <25, hasil kesesuaian positif dan negatif adalah 100%. Nilai Ct < 30 hasil kesesuaian positif 85,3 % dan negatif adalah 100%. Nilai Ct < 40 , hasil kesesuaian positif 51,1 % dan negatif adalah 100%.  Kesimpulan : Swab bukal dapat digunakan sebagai pemeriksaan alternatif pada pemeriksaan SARS- CoV-2. ......Background: COVID-19 caused by the SARS-COV-2 virus. WHO published protocol for the detection of the virus using the real time RT-PCR from nasopharyngeal and oropharynx swab specimens. This method is invasive. Required an examination technique that is relatively safe and comfortable. This study aims to see the effectiveness of the buccal swab as an alternative to the SARS-CoV-2 examination. Methods: This diagnostic test study from 2020 to 2021, specimens of nasopharyngeal, oropharyngeal and buccal swabs from COVID-19. Specimens underwent an optimization, viral RNA extraction and real time RT-PCR. Result : This study collected 68 specimens from COVID- 19 patients. The results of positive nasopharyngeal, oropharynx and buccal tests were 24 specimens. The results of a positive nasopharynx and oropharynx test with a negative buccal test were 23 specimens. Based on the values ​​of Ct < 20 and Ct < 25 , the results of positive agreement and negative are 100%. The value of Ct < 30 and Ct < 40  results in a positive agreement are 85.3% and 51,1 %. The negative  results are 100%. Conclusion : Buccal swab can be used as an alternative test for SARS-CoV-2 examination.

Abstrak :
Rotavirus causes 25?55% of all hospital admissions for diarrhea and approximately 611.000 deaths every year in developing countries. Clinically, it is not possible to recognize the diarrhea caused by rotavirus and other infections. To know a causative agent of rotavirus gastroenteritis, availability of an accurate diagnosis assay is necessary. Therefore, we developed real time RT-PCR assay (rRT-PCR) assay for confirmation of infections of Group A or C rotaviruses simultaneously. A total of 54 stool samples obtained from pediatric patients (< 5 years old) was used in this study. All samples were tested for Group A rotavirus by Serological rapid test. Result of serological rapid test was compared with rRT-PCR assay to obtain the test accuracies of both assays. Result of this study showed that rates of positive testing for Group A rotavirus by serological rapid test and the rRT-PCR assay were 22.22% and 18.50%, espectively. Forty-two serology-negative specimens for Group A rotavirus were also PCR negative (100% specificity). Two serology-positive specimens for Group A rotavirus was rRT-PCR negative (confirmed by electrophoresis gel); therefore, rRT-PCR assay represents the decrease of 3.70% in the number of specimens that are positive for Group A rotavirus. For Group C rotavirus, all tested samples were no rRT-PCR positive and the results need to be confirmed in the future.

Abstrak :
Demam akut yang disertai gejala mirip demam Dengue nyeri kepala, nyeri sendi, ruam, perdarahan perifer seperti mimisan, ptekie adalah gejala yang paling sering dikeluhkan oleh pasien. Gejala-gejala tersebut seringkali diakibatkan infeksi arbovirus yang sangat endemik di Indonesia sebagai negara tropis. Deteksi agen penyebab infeksi tersebut sangat diperlukan untuk penatalaksanaan yang tepat. Studi ini melakukan uji optimasi untuk deteksi molekuler virus penyebab demam mirip demam Dengue, meliputi DENV, ZIKV, WNV, JEV, YFV, CHIKV, dan Hantavirus menggunakan RT PCR. Primer pada studi ini dirancang menggunakan perangkat lunak online Primer-BLAST dari NCBI dan primer-primer tersebut memenuhi kriteria untuk reaksi RT PCR. Kontrol positif pada real time RT-PCR menggunakan DNA sintetik yang dirancang sesuai dengan amplicon target virus. DNA sintetik sepanjang 1.047 pasang basa dirancang untuk digunakan pada virus ZIKV, JEV, YFV, WNV, CHIKV, dan Hantavirus. Suhu penempelan optimum pada primer-primer adalah 600C kecuali primer flavivirus universal yaitu 560C. Limit deteksi primer JEV mencapai 4.355 salinan DNA setiap reaksi real time RT PCR. Tidak terdapat reaksi silang maupun positif palsu pada sampel RNA DENV serotipe 2 maupun pada sampel orang sehat yang digunakan pada studi ini. Sebagai kesimpulan, studi ini menghasilkan primer dan protokol real time RT-PCR yang berpotensi untuk dikembangkan lebih lanjut untuk digunakan dalam uji diagnostik pada sampel pasien demam akut menyerupai gejala demam Dengue.

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Acute fever with Dengue like fever symptoms headache, rash, joint pain, perifer bleeding like ptechie, rinhorrhea is general symptoms that often being complained by patient. Usually the etiology agent of the symptoms are arbovirals which are endemic in Indonesia as a tropical country. In this case, molecular detection is very important to confirm the etiology of disease for prompt and adequate management. This study optimized viral molecular detection as an etiology agent for Dengue like fever symptoms using real time RT PCR. The viruses that were investigated were DENV, ZIKV, WNV, JEV, YFV, CHIKV, and Hantavirus. Primer were designed used Primer BLAST software from NCBI. Those primers fulfilled the good primer requirements and could be used in real time RT PCR reaction. Synthetic DNA with 1.047 base pairs was designed based on amplicon target to be used as control positive for ZIKV, JEV, YFV, WNV, CHIKV, and Hantavirus. The optimal annealing temperature for all primers were at 600C except for flavivirus universal primer was at 560C. The limit of detection of JEV primer was 4355 copies DNA per reaction. Cross reactivity between all primers with DENV serotype 2 RNA and healthy person sample were not found. This study still need RNA viruses as negative or positive control and clinical sample to determine the sensitivity and specificity. As a conclusion, this study provided primers and real time RT PCR protocol that potentially be further developed as diagnostic tools for patient with Dengue like fever symptoms.