Tesis

Uji Diagnostik Kit BioCoV-19 RT-PCR Dengan N1N2 CDC Untuk Mendeteksi SARS-CoV-2 = Diagnostic Test of BioCov-19 RT-PCR Kit With N1N2 CDC For Detecting SARS-CoV-2.

Latar Belakang: Pada bulan Desember, 2019, serangkaian kasus pneumonia dengan penyebab yang tidak diketahui muncul di Wuhan, Hubei, Cina, dengan presentasi klinis sangat menyerupai virus pneumonia. Analisis data menunjukkan adanya coronavirus baru, yang diberi nama SARS-CoV-2. WHO sebagai lembaga kesehatan dunia telah menyatakan coronavirus disease-2019 (COVID-19) sebagai pandemi yang mengancam lebih dari 114 negara. Berdasarkan WHO dan CDC saat ini pemeriksaan menggunakan metode uji molekular merupakan pemeriksaan yang disarankan, yaitu real time PCR, saat ini terdapat banyak kit untuk mendeteksi COVID-19, salah satu nya adalah BioCoV-19 RT-PCR kit. Metode: Uji diagnostik dengan metode potong lintang dilakukan pada bulan Oktober hingga Desember 2020 dengan subjek pasien yang datang secara mandiri ke Laboratorium Mikrobiologi Klinik Fakultas Kedokteran Universitas Indonesia yang sesuai dengan kriteria inklusi dan eksklusi. Subjek penelitian dilakukan pengambilan spesimen dari nasofaring dan orofaring kemudian spesimen di ekstraksi dan dilakukan pengujian menggunakan Kit BioCoV-19 RT-PCR dengan N1N2 CDC, kemudian dilakukan uji minimal deteksi dari Kit BioCoV-19 RT-PCR dan N1N2 CDC untuk menilai sensitivitas analisis. Untuk melihat potensi reaksi silang dilakukan uji dengan 12 bakteri, tujuh virus dan tiga jamur. Karakteristik subjek penelitian di jelaskan secara deskriptif. Hasil Penelitian: Didapatkan 135 spesimen dengan hasil uji 80 spesimen positif dari uji N1N2 CDC dan 78 dari uji Kit BioCoV-19 RT-PCR, hasil uji negative didapatkan 55 spesimen baik dari uji Kit BioCoV-19 RT-PCR maupun N1N2 CDC. Hasil sensitivitas dan spesifisitas BioCoV-19 RT-PCR Kit secara umum adalah 97,50% dan 100%, dengan Nilai Duga Positif (NDP) 100% dan Nilai Duga Negatif (NDN) 96,49%. Hasil uji minimal deteksi (LOD) untuk primer-probe N1N2 CDC dan BioCoV-19 RT-PCR Kit setelah dilakukan dilusi bertingat sebanyak enam kali pengenceran yakni 3,5 kopi/reaksi (rerata nilai Ct 35,21). Uji reaksi silang tidak terdeteksi adanya reaksi silang dari 12 bakteri, tujuh virus dan tiga jamur. Karakteristik subjek penelitian lebih banyak pada laki-laki sebanyak (61,5%), untuk usia lebih banyak pada usia berkisar 20-40 tahun (56,29%), gejala klinis pasien saat datang lebih banyak gejala ringan. Kesimpulan: BioCov-19 RT-PCR Kit dapat digunakan sebagai alternatif untuk mendeteksi SARS-CoV-2. Sensitivitas dan spesifisitas BioCoV-19 RT-PCR Kit adalah 97,5% dan 100% Sensitivitas berdasarkan Ct cukup baik, namun sensitivitas untuk Ct ≥ 35 sedikit lebih rendah. Karakteristik subjek penelitian Sebagian besar adalah laki-laki (61,5%), usia lebih banyak pada rentang 20-40 tahun (56,29%), sebagian besar mengalami gejala.
Kata Kunci: SARS-CoV-2, RT-PCR, COVID-19, deteksi, N1N2 CDC, BioCoV-19 RT-PCR


Background: In December, 2019, a series of pneumonia cases of unknown cause appeared in Wuhan, Hubei, China, with a clinical presentation strongly resembling viral pneumonia. Analysis of the data indicated the presence of a new coronavirus, which was named SARS-CoV-2. WHO as a world health institution has declared coronavirus disease-2019 (COVID-19) a pandemic that threatens more than 114 countries. According to the WHO and the CDC, currently testing using the molecular test method is the recommended examination, namely real time PCR. Currently there are many kits to detect COVID-19, one of which is the BioCoV-19 RT-PCR kit. Method: Diagnostic tests using the cross-sectional method were carried out from October to December 2020 with patient subjects who came independently to the Laboratorium Mikrobiologi Klinik FKUI who met the inclusion and exclusion criteria. The research subjects collected specimens from the nasopharynx and oropharynx, then the specimens were extracted and tested using the BioCoV-19 RT-PCR Kit and N1N2 CDC, then a minimal detection test was carried out from the BioCoV-19 RT-PCR Kit and N1N2 CDC to assess the sensitivity of the analysis. To see the potential for cross reactions, tests were carried out with 12 bacteria, seven viruses and three fungi. The characteristics of the research subjects are described descriptively. Results: Among 135 specimens were obtained with positive test results, 80 specimens from the CDC N1N2 test and 78 from the BioCoV-19 RT-PCR Kit test, negative test results obtained 55 specimens from both the BioCoV-19 RT-PCR Kit and the CDC N1N2 test. The sensitivity and specificity results of the BioCoV-19 RT-PCR Kit in general were 97.50% and 100%, with a positive predictive value (NDP) of 100% and a negative predictive value (NDN) of 96.49%. Minimum detection (LOD) test results for the N1N2 CDC and the BioCoV-19 RT-PCR Kit after six dilutions of 3.5 copies/reaction (mean Ct value 35.21). The cross-reaction test did not detect any crossreaction of 12 bacteria, seven viruses and three fungi. The characteristics of the study subjects were mostly male (61.5%), for ages ranging from 20-40 years (56.29%), the clinical symptoms of the patients when they arrived were more mild symptoms. Conclusion: The BioCov-19 RT-PCR Kit can be used as an alternative to detect SARSCoV-2. The sensitivity and specificity of the BioCoV-19 RT-PCR Kit are 97.5% and 100%. The sensitivity based on Ct is quite good, but the sensitivity for Ct ≥ 35 is slightly lower. Characteristics of research subjects most were men (61.5%), mostly in the age range of 20-40 years (56.29%), most of them experienced light symptoms.
Keywords: SARS-CoV-2, RT-PCR, COVID-19, detection, N1N2 CDC, BioCoV-19 RT-PCR