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Abstrak :
Virus dengue (DENV) adalah penyebab penyakit infeksi yang endemik di 100 negara di dunia. DENV dapat menyebabkan infeksi primer dan sekunder. Infeksi sekunder diperkirakan lebih berat dan akan menyebabkan menjadi Demam Berdarah Dengue (DBD) dan Sindrom Renjatan Dengue (SRD). Penatalaksanaan yang lebih dini dan tepat akan membantu mengurangi terjadinya kasus berat seperti DBD dan SRD. Teknik diagnostik yang dikembangkan belum ada yang dapat mendeteksi secara cepat dan tepat pada awal infeksi terutama untuk virus dengue strain Indonesia. Tujuan penelitian ini untuk mendapatkan kondisi yang optimal untuk deteksi DENV dan analisis validasi tehnik in house multiplek realtime Reverse Transcriptase-Polimerase Chain Reaction (rRT-PCR) sehingga dapat digunakan untuk deteksi dini dan cepat infeksi DENV. Rancangan penelitian ini adalah penelitian eksperimental laboratorium. Strain standar DENV diisolasi dengan kit Roche® dan spesimen diekstraksi dengan kit Qiagen®. Strain standar DENV digunakan untuk optimasi suhu annealing dan konsentrasi primer masing-masing serotipe DENV dengan metode in house multiplek rRT-PCR berbasis SYBR green. Sebagai pembanding digunakan RT-PCR konvensional dengan menggunakan primer di daerah C-PrM. Primer in house multiplek rRT-PCR didesain di daerah envelope pada masing masing serotipe. Analisis limit of detection (LOD) dilakukan dengan pengenceran titer virus 105, 104, 103, 102, 10 dan 1 FFU/ml (in house multiplek rRT-PCR) pada keempat serotipe. Hasil in house multiplek rRT-PCR dibandingkan dengan hasil RT-PCR konvensional Lanciotti pada pasien dengue yang telah masuk dalam kriteria inklusi dan eksklusi. Suhu annealing optimal didapatkan pada suhu 58oC sedangkan konsentrasi optimal masing-masing primer untuk in house multiplek rRT-PCR adalah 4 pmol. LOD RNA pada DENV-1, DENV-2, DENV-3, DENV-4 adalah 1 FFU/ml, 10 FFU/ml, 104 FFU/ml dan 1 FFU/ml. In house multiplek rRT-PCR dibandingkan RT-PCR konvensional mempunyai sensitivitas sebesar 100%, spesifisitas 94,2 %, nilai prediksi positif 85,7% dan nilai prediksi negatif 100 %. In house multiplek rRT-PCR berbasis SYBR green merupakan metode yang dini, cepat dan tepat untuk deteksi DENV pada awal infeksi dengan sensitivitas dan spesifisitas yang baik sebagai metode diagnostik infeksi dengue dimasa mendatang. ...... Dengue virus (DENV) is an infectious disease that is endemic in 100 countries in the World. Dengue virus infections can cause primary and secondary. Secondary infection is estimated to be more severe DHF and SDD. Developed diagnostic technique that no one has been able to quickly and accurately detect the infection early, especially for the Indonesian strain of dengue virus. The purpose of this study is to obtain optimal conditions for the detection of dengue infection and analysis techniques in-house validation of multiplex real-time Reverse Transcriptase-Polymerase Chain Reaction (rRT-PCR) for the Indonesian strain of dengue virus. Design of this study is an experimental research laboratory. Standard strains of dengue virus was isolated with a kit Roche® and the specimen was extracted with Qiagen® kit. Standard strains of dengue virus is used for optimization primer annealing temperature and the concentration primers of each serotype dengue virus by multiplex rRT-PCR method based on SYBR green. Primers for RT-PCR conventional based lanciotii et al while rRT-PCR primer was designed in the envelope gen at each serotype. Limit of detection (LOD) by diluting the virus titer 105, 104, 103, 102 FFU /ml performed by rRTPCR in all four serotypes. The results of multiplex rRT-PCR compared with results of conventional RT-PCR in patients with dengue lanciotti superbly into the criteria inclusion and exclusion. Optimal annealing temperature results obtained at a temperature of 58oC and optimal primer concentration of 4 pmol of each primer for 25 ul total reaction. LOD RNA in DENV-1, DENV-2, DENV-3 and DENV-4 at titer of 1 FFU / ml, 10 FFU/ml, 104 FFU/ml and 1 FFU/ml. .In house multiplex rRT-PCR compared with RT-PCR has a sensitivity of 100%, specificity 95,2%, positive predictive value 85,7% and negative predictive value of 100%. In-house multiplex rRT-PCR with SYBR green-based research is a method that is rapid and precise detection of dengue virus in early infection with good sensitivity and specificity compared to RT-PCR as a diagnostic method in the future dengue infection.

Abstrak :
Latar Belakang. Saat ini sedang terjadi pandemi COVID-19 di seluruh dunia, pandemi ini dimulai dari Wuhan, Cina. Virus SARS-CoV-2 penyebab COVID-19 sangat menular dan penyebarannya sangat cepat, sehingga memerlukan penanganan khusus seperti isolasi atau karantina. Gejala penyakit COVID-19 menyerupai gejala infeksi saluran pernapasan akut yang disebabkan oleh patogen lain seperti SARS, influenza, rhinovirus, dll. Diagnosis penyakit COVID-19 perlu ditentukan untuk membedakan dari ISPA yang diakibatkan oleh patogen lain. Beberapa uji diagnostik telah di kembangkan untuk deteksi cepat penyebab COVID-19. Tujuan. Tujuan penelitian ini adalah membandingkan alat diagnostik kit antigen ”Genbody COVID-19 Test Ag®” dengan rRT-PCR yang menjadi refensi test, untuk mendapatkan alat diagnostik yang murah, cepat dan akurat serta memiliki kemampuan yang setara dengan rRT-PCR. Metode. Penelitian ini merupakan uji banding dengan desain penelitian potong lintang dan metode pengumpulan spesimen secara consecutive sampling pada pasien contact tracing COVID-19. Penelitian dilakukan di Fasilitas Pelayanan Kesehatan (FASYANKES) Puskesmas Kecamatan Tanah Abang, Sawah Besar, Senen dan Laboratorium Mikrobiologi Klinik (LMK) FKUI pada bulan Oktober 2020-Desember 2020. Sampel penelitian merupakan swab Nasofaring dan oropharing dari pasien contact tracing COVID-19 yang dilakukan pemeriksaan rRT-PCR menggunakan reagen LiliF COVID-19 Real Time PCR kit dan pemeriksaan antigen menggunakan “Genbody COVID-19 Test Ag®”. Analisis penelitian ini menggunakan tabel 2x2. Hasil. Dari 233 sampel sebanyak 80 (34,33%) sampel positif rRT-PCR dan 53 (22,74%) sampel yang positif pada kit antigen. Kit antigen yang digunakan pada penelitian ini mempunyai sensitivitas 66,25% (55,89-76,61), spesifisitas 100% (100-100), Nilai Duga Positif (NDP) 100% (100-100), Nilai Duga Negatif (NDN) 85% (79,78-90,22) dan akurasi 88,41%. Pada hasil rRT-PCR dengan CT < 20, kit test antigen mempunyai sensitivitas 97,14% (91,62-102,66), spesifisitas 100% (100-100) dan pada CT ≥ 21-≥ 30 sensitivitas kit antigen terus menurun. Kesimpulan. Pemeriksaan COVID-19 menggunakan kit test antigen “Genbody COVID-19 Test Ag®” mempunyai sensitivitas rendah yang tidak sesuai dengan rekomendasi WHO. Kit antigen ini mempunya sensitivitas yang tinggi pada sampel dengan hasil rRT-PCR pada CT rendah (CT <20). ......Introduction. Currently, the COVID-19 pandemic is happening over the world, this pandemic started in Wuhan, China. The SARS-CoV-2 virus that causes COVID-19 is highly contagious, spreads very quickly and requiring special handling such as isolation or quarantine. The symptoms of COVID-19 resemble an acute respiratory infection caused by other pathogens such as SARS, influenza, rhinovirus, etc. The diagnosis of COVID-19 disease needs to be determined to distinguish it from acute respiratory infection caused by other pathogens. Several diagnostic tests have been developed for rapid detection of the cause of COVID-19. Aim. The research aims to compare the antigen kit "Genbody COVID-19 Test Ag®" with rRT-PCR as a reference test to obtain an affordable, fast and accurate diagnostic tool and have equivalent capabilities as rRT-PCR. Method. The research is a comparative testing with a cross-sectional design and consecutive sampling method for collecting specimens in COVID-19 contact tracing patients. The research was conducted at the Health Service Facility (FASYANKES) Puskesmas Kecamatan Tanah Abang, Sawah Besar, Senen and Laboratorium Mikrobiologi Klinik (LMK) FKUI in October 2020-December 2020. The research samples were nasopharyngeal and oropharyngeal swabs from COVID-19 contact tracing patients who were carried out rRT-PCR test using LiliF COVID-19 Real Time PCR kit and antigen test using “Genbody COVID-19 Test Ag®”. The research analysis used a 2x2 table. Results. Of the 233 samples, 80 (34.33%) were positive for rRT-PCR and only 53 (22.74%) were positive for the antigen kit. The antigen kit used in this research had a sensitivity of 66.25% (55.89-76.61), specificity 100% (100-100), Positive Prediction Value (NDP) 100% (100-100), Negative Suggestion Value ( NDN) 85% (79.78-90.22) and accuracy 88.41%. On the results of rRT-PCR with CT < 20, the antigen test kit had a sensitivity of 97.14% (91.62-102.66), specificity 100% (100-100) and at CT 21-30 the sensitivity of the antigen kit continued to decrease. Summary. The COVID-19 examination using the “Genbody COVID-19 Test Ag®” antigen test kit has a low sensitivity which is not in accordance with WHO recommendations. This antigen kit has high sensitivity in rRT-PCR results at CT (CT < 20).

Abstrak :
Latar Belakang. SARS-CoV-2 sebagai penyebab COVID-19 pertama kali terdeteksi pada sampel klaster pasien di Provinsi Hubei, China pada Desember 2019. Pada mulanya klaster pasien tersebut memiliki gejala seperti demam, batuk, sesak nafas, dan gejala lainnya yang tidak spesifik. Alat uji Rapid Antigen Test (RAT) dapat dijadikan alternatif untuk diagnosis klinis COVID-19. Tujuan. Penelitian ini bertujuan untuk mendapatkan rekomendasi mengenai alternatif spesimen dan metode deteksi SARS-CoV-2. Metode. Desain penelitian ini merupakan uji diagnostik studi potong lintang dengan pengumpulan spesimen secara consecutive sampling. Subjek penelitian yaitu pasien yang memiliki kontak dengan kasus infeksi SARS-CoV-2 yang terkonfirmasi dengan atau tanpa gejala klinis COVID-19 di Fasilitas Pelayanan Kesehatan (Fasyankes) dan Laboratorium Mikrobiologi Klinik (LMK) FKUI dengan jumlah sampel 221. Analisis data dengan tabulasi silang dan perhitungan sensitivitas, spesifisitas, PPV, dan NPV. Hasil. Deteksi antigen menggunakan spesimen nasal memiliki nilai sensitivitas 32,35%, spesifisitas 99,35%, PPV 95,65%, NPV 76,77%, akurasi 78,73%. Tingkat positifitas pada spesimen nasofaring 34,84%, spesimen orofaring 30,32%, dan nasal 30,77%. Kesimpulan. Hasil uji rRT-PCR pada beberapa jenis spesimen menunjukkan bahwa spesimen nasal dan orofaring dapat dijadikan pilihan selain spesimen nasofaring. Penggunaan kit deteksi antigen dapat dilakukan untuk pelacakan kontak COVID-19 atau untuk diagnosis, terutama untuk daerah yang memiliki keterbatasan akses diagnosis menggunakan rRT-PCR. ......Introduction. The SARS-CoV-2 as the cause of COVID-19 was first detected in a cluster sample of patients in Hubei Province, China in December 2019. The first patient had symptoms such as fever, cough, shortness of breath, and other non-specific symptoms. Rapid Antigen Test can be used as an alternative for diagnosis of COVID-19. Aim. This study aims to obtain recommendations alternative specimens and detection methods for SARS-CoV-2. Method. The design of this study is a cross-sectional diagnostic test with consecutive sampling. The research subjects were patients who had contact with confirmed cases of SARS-CoV-2 infection with or without clinical symptoms of COVID-19 at Health Service Facilities (Fasyankes) and Laboratorium Mikrobiologi Klinik (LMK) FKUI with a total sample of 221. Data analysis using cross tabulation to calculate the sensitivity, specificity, PPV, and NPV. Results. The positivity rate for nasopharyngeal specimens was 34.84%, oropharyngeal specimens 30.32%, and nasal specimens 30.77%. Antigen detection using nasal specimens has sensitivity 32.35%, specificity 99.35%, PPV 95.65%, NPV 76.77%, accuracy 78.73%. Conclusion. The results of the rRT-PCR test on several types of specimens indicate that nasal and oropharynx specimens can be used as an alternative to nasopharyngeal specimens. The use of antigen detection kits can be carried out for COVID-19 contact tracing or for diagnosis, especially for areas that have limited access to diagnosis using rRT-PCR.

Abstrak :
Latar Belakang. SARS-CoV-2 menimbulkan beban yang sangat besar pada masyarakat, sistem ekonomi dan kesehatan di seluruh dunia. Berbagai langkah diambil untuk mengendalikan penyebarannya. Langkah – langkah tergantung pada diagnosis yang tepat waktu dan akurat dari orang yang terinfeksi virus. Penyebaran COVID-19 yang cepat, deteksi virus yang cepat dan tepat memegang peranan penting untuk mengendalikan infeksi dan membantu pasien untuk mencegah perkembangan penyakit lebih lanjut. Metode real-time reverse transcriptase-PCR (rRT-PCR) sangat dianjurkan untuk deteksi SARS-CoV-2 sebagai uji kualitatif spesifik dan sederhana. Selain metode menggunakan rRT-PCR, telah terdapat metode dengan pendekatan yang berbeda untuk mendeteksi SARS-CoV-2. Salah satu metode tersebut adalah VereCoVTM yang memiliki cara kerja berdasarkan teknik PCR dan hibridisasi, namun performa dari VereCoVTM masih belum diketahui. Tujuan. Penelitian bertujuan untuk mengetahui kesesuaian dan kaitannya dengan gejala klinis menggunakan uji VereCoVTM dengan rRT-PCR dalam mendeteksi SARS-CoV-2. Metode. Penelitian ini menggunakan consecutive sampling. Sampel penelitian yaitu sampel swab nasofaring dan orofaring yang diambil kemudian dianalisis di Laboratorium Mikrobiologi Klinik Fakultas Kedokteran Universitas Indonesia. Analisis data menggunakan Kappa Cohen. Selanjutnya, dilakukan analisis data kesesuaian hasil pemeriksaan kedua uji pada pasien dengan gejala dan pasien tanpa gejala. Hasil. Perbedaan positivity rate antara VereCoVTM dan rRT-PCR sebesar 68% menunjukkan bahwa VereCoV™ memberikan hasil negatif palsu. Jika disandingkan terhadap data hasil rRT-PCR, maka ambang batas deteksi VereCoVTM setara dengan nilai Ct<32 atau setara dengan 118 salinan RNA. Tidak dijumpai reaksi silang dengan beberapa mikroorganise. Konsistensi antara kedua uji ditunjukan oleh koefisien Kappa (=0,609). Hal ini menggambarkan kesesuian yang moderate atau sedang. Analisis lebih lanjut menunjukkan bahwa kesesuaian kedua pemeriksaan meningkat pada pasien yang bergejala (90%), sebaliknya menurun pada pasien tanpa gejala (76,5%). VereCoVTM lebih sesuai digunakan untuk mendeteksi SARS-CoV-2 pada pasien yang bergejala. Hasil negatif pada VereCoVTM tidak dapat mengesampingkan kemungkinan infeksi COVID-19, sehingga perlu pemeriksaan lebih lanjut untuk mendukung diagnosis yang tepat. Kesimpulan. VereCoV™ lebih cocok untuk mendeteksi SARS-Cov-2 pada pasien yang bergejala. VereCoVTM tidak menunjukan adanya reaksi silang dengan beberapa mikroorganisme, yang terdapat pada saluran pernapasan. ......Background. SARS-CoV-2 places an enormous burden on society, economic systems and health around the world. Various steps were taken to control its spread. These steps depend on timely and accurate diagnosis of the person infected with the virus. The rapid spread of COVID-19, rapid and precise detection of the virus play an important role in controlling infection and helping patients to prevent further disease progression. The real-time reverse transcriptase-PCR (rRT-PCR) method is highly recommended for the detection of SARS-CoV-2 as a simple and specific qualitative test. In addition to the method using rRT-PCR, there have been methods with different approaches to detect SARS-CoV-2. One of these methods is VereCoVTM which has a working method based on PCR and hybridization techniques, but the performance of VereCoVTM is still unknown. Aim. The aim of the study was to determine the suitability and relation to clinical symptoms using the VereCoVTM test with rRT-PCR in detecting SARS-CoV-2. Method. This study used consecutive sampling. The samples of the study was swab samples of the nasopharynx and oropharynx which were taken and analyzed at the Clinical Microbiology Laboratory, Faculty of Medicine, Universitas Indonesia. Data was then analyze using Kappa Cohen. Furthermore, the data analysis of the suitability of the results of the two tests was carried out in patients with symptoms and patients without symptoms. Results. The difference in positivity rate between VereCoVTM and rRT-PCR of 68% indicates that VereCoV™ gives a false negative result. When compared to the rRT-PCR data, the VereCoVTM detection threshold is equivalent to a Ct value <32 or equivalent to 118 copies of RNA. No cross reactions were found with some microorganisms. Consistency between the two tests is indicated by the Kappa coefficient (= 0.609). This describes a moderate or moderate fit. Further analysis showed that the concordance of the two examinations increased in symptomatic patients (90%), on the contrary decreased in asymptomatic patients (76.5%). VereCoVTM is more suitable for detecting SARS-CoV-2 in symptomatic patients. A negative result on VereCoVTM cannot rule out the possibility of COVID-19 infection, so further tests are needed to support a correct diagnosis. Summary. VereCoV™ is more suitable for detecting SARS-Cov-2 in symptomatic patients. VereCoVTM did not show any cross-reactivity with some microorganisms, which are present in the respiratory tract.

Abstrak :
Background: real-time RT-PCR was recommended by WHO for COVID-19 diagnosis. The cycle threshold (Ct) values were expected to have an association with clinical manifestation. However, the diagnostic modalities such as quantitative molecular detection and virus isolation were not yet available for the routine test. This study has been conducted to analyze the relationship between the Ct values of qualitative rRT-PCR and the clinical manifestation and to describe the factors determining the result. Methods: from March to April 2020, specimens were sent to our laboratory from different healthcare centers in Jakarta. The patient's characteristic and clinical manifestation were extracted from the specimen's epidemiology forms. The specimens extracted and tested using rRT-PCR, and the Ct value were collected. The data were analyzed using the appropriate statistic test. Results: from 339 positive results, the mild to moderate case was 176 (52%) and the severe cases was 163 (48%). Female was dominant in the mild to moderate cases (58%), while the male was prevalent in the severe cases (60%). The median age for mild to moderate case was 35 years old and severe cases was 49 years old. Statistical analysis found relationship between both group with gender (p = 0.001) and age (p < 0.001), but not with the Ct value. Conclusion: many variables in specimen sampling and processing could affect the Ct value result. In addition, the disease's severity was depended with the host immune response, regardless the number of virus. There was suggested no significant difference between the Ct values of mild-moderate and severe COVID-19, and thus should not be loosely interpreted.