Pemeriksaan Laboratorium COVID-19 (2019-nCoV) Irmawanty Setiaputri RS PARU dr.H.A ROTINSULU WHO Novel Corona Virus= 2019-nCoV COVID-19 Corona Virus Disease Karakteristik COVID 19 (2019-nCoV) • Virus corona adalah zoonosis (ditularkan dari hewan dan manusia) • Merupakan virus single-strand, positivesense RNA, panjang genom bervariasi dari 26 to 32 kb • Transmisi virus melalui droplet namun diduga dapat melalui airborne Guidance: 1 Biosafety Level 2 Lab Guidance 5 6 Specimen Type 3 Safety Procedures During Sample Collection and Transport 4 Testing of 2019-nCoV in Reference Laboratories Reporting of Cases and Test Results Diagnostic Knowledge Gaps SPESIMEN UNTUK PENEGAKAN DIAGNOSIS Jenis spesimen dan prioritas Untuk meningkatkan deteksi CDC merekomendasikan: •Pengambilan 3 jenis spesimen dari : • saluran nafas bagian atas • saluran nafas bagian bawah • Serum darah •Jika memungkinkan/diperlukan: urine dan feses Spesimen Jumlah Container Saluran nafas bagian atas - Apus nasofaring - Apus orofaring 1 apus 1 apus Virus Transport Media Saluran nafas bagian bawah - Sputum - Tracheal aspirate - BAL 2-3 ml 2-3 ml Pot sputum Sputum trap Darah - Serum Whole blood Tabung eppendorf 5 ml Specimen Type Tipe Spesimen CDC, WHO 2020 Nasopharyngeal swab: •Masukkan swab kedalam lubang hidung sejajar langit langit mulut. •Diamkan beberapa saat agar sekret NP meresap pada swab, lakukan dari kedua belah lubang hidung •Segera masukan swab ke dalam tabung berisi VTM Oropharyngeal swab (e.g., throat swab): •Lakukan swab pada daerah belakang faring jangan sampai menyentuh lidah •Segera masukkan swab ke dalam tabung berisi VTM Viral Transport Medium (VTM) Eppendorf tube Swab sintetik berbahan DACRON dgn GAGANG PLASTIK !!!! Jangan pakai swab calcium alginate swabs atau swab gagang kayu : karena dapat mengandung bahan yang menginaktivasi virus dan menghambat proses PCR Masukkan SEGERA swab O/N disatukan ke tabung VTM Simpan spesimen pada suhu 4°C segera kirim ke reff lab dalam rantai dingin Safety procedures sample transport Safety procedures • Triple layer packaging Form Pengantar Spesimen Balitbangkes • Patients that meet the case definition for suspected 2019nCoV should be screened for the virus with PCR. • Laboratories may desire to use a pan-coronavirus assay for amplification followed by sequencing of amplicons from non- conserved regions for characterization and confirmation. • Several molecular assay have been and are currently under development. Some groups shared their molecular assays, most procedures assume a basic familiarity with PCR/RT-PCR assay. Testing 2019-nCoV Testing of 2019-nCoV in reference laboratories Pemeriksaan COVID 19 di BALITBANGKES Menggunakan 2 protokol dari: 1. US CDC: rRT-PCR 2. Jerman: Menggunakan 2 target gen: - Gen N (nucleocapsid) - gen RdRp (RNA-dependent RNA polymorphism) • TAT: + 2 hari In-house developed molecular assays https://www.who.int/emergencies/diseases/novel- coronavirus2019/technical-guidance/laboratory-guidance • China CDC Primers and probes for detection 2019-nCoV (24 January 2020) • Diagnostic detection of Wuhan coronavirus 2019 by real-time RT-PCR – Charité, Berlin Germany (17 January 2020) • Detection of 2019 novel coronavirus (2019-nCoV) in suspected human cases by RT-PCR – Hong Kong University (23 January 2020) • PCR and sequencing protocol for 2019-nCoV - Department of Medical Sciences, Ministry of Public Health, Thailand (Updated 28 January 2020) • PCR and sequencing protocols for 2019-nCoV- National Institute of Infectious Diseases Japan (24 January 2020) • US CDC panel primer and probes– U.S. CDC, USAV – U.S. CDC, USA (28 January 2020) https://www.who.int/emergencies/diseases/novel- coronavirus2019/technical-guidance/laboratory-guidance Testing 2019-nCoV In-house developed molecular assays Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China Cases Characteristic Chest X-Ray • •Usia 21 – 82 (55.5) •man : woman = 3 : 1 •51% w chronic ilnness: •CV dan CVD 40% •ICU: ARDS (17%), acute respiratory injury (8%), septic shock (4%) •Co-infection: – Bacteria (1%) – Fungus (4%) – Other virus (0%) Chen N, et al. Published Online January 29, 2020 Bilateral pneumonia (75%), unilateral pneumonia (25%), groundglass opac (14%) Blood Tests • • • • • • • • • • • Leukocytosis (24%) Lymphopenia (35%) Anemia (51%) Prothrombin time decreased (30%) D-dimer increase (36%) Albuminemia (98%) ALT increased (35%) LDH increased (76%) ESR increased (85%) Serum feritin increased (63%) CRP increased (86%) Hal yang perlu diperhatikan dalam Pengambilan spesimen Perhatikan kewaspadaan standard dan Kewaspadaan berbasis transmisi • Kebersihan tangan • Gunakan APD lengkap a. Masker N95/respirator partikulat b. Sarung tangan c. Pelindung wajah atau mata/face shield d. Baju/coat/gaun/apron e. Tutup kepala f. Sepatu tertutup atau memakai cover shoes • Higiene petugas • Kebersihan ruangan: dekontaminasi permukaan dengan desinfektan TERIMA KASIH
