Irmawanty Setiaputri

RS PARU dr.H.A ROTINSULU

Pemeriksaan LaboratoriumCOVID-19

(2019-nCoV)

WHONovel Corona Virus= 2019-nCoV

COVID-19Corona Virus Disease

Karakteristik COVID 19 (2019-nCoV)

• Virus corona adalah zoonosis (ditularkan dari hewan dan manusia)

• Merupakan virus single-strand, positive-sense RNA, panjang genom bervariasi dari 26 to 32 kb

• Transmisi virus melalui droplet namun diduga dapat melalui airborne

Guidance:

1

2

3

4

6

Specimen Type

Biosafety Level

Safety Procedures During Sample Collection and Transport

Lab

GuidanceTesting of 2019-nCoV in Reference Laboratories

Diagnostic Knowledge Gaps

Reporting of Cases and Test Results5

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 UNTUK PENEGAKAN DIAGNOSIS

Spe

cim

en

Typ

e

Tipe Spesimen

Spesimen Jumlah Container

Saluran nafas bagian atas

- Apus nasofaring- Apus orofaring

1 apus1 apus

Virus Transport Media

Saluran nafas bagian bawah

- Sputum- Tracheal aspirate- BAL

2-3 ml2-3 ml

Pot sputumSputum trap

Darah - Serum Whole blood 5 ml

Tabung eppendorf

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

Safe

typ

roce

du

res

Safety procedures sample transport

• Triple layer packaging

Form Pengantar Spesimen Balitbangkes

Test

ing

20

19

-nC

oV

• Patients that meet the case definition for suspected 2019-nCoV 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 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

https://www.who.int/emergencies/diseases/novel- coronavirus-2019/technical-guidance/laboratory-guidance

In-house developed molecular assays

Test

ing

20

19

-nC

oV

https://www.who.int/emergencies/diseases/novel- coronavirus-2019/technical-guidance/laboratory-guidance

In-house developed molecular assays

• 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)

Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China

Chest X-Ray

Blood Tests

• Bilateral pneumonia (75%), unilateral pneumonia (25%), ground-glass opac (14%)

• 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%)

Cases Characteristic

•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

Hal yang perlu diperhatikan dalam Pengambilan spesimen

Perhatikan kewaspadaan standard dan Kewaspadaan berbasis transmisi

• Kebersihan tangan

• Gunakan APD lengkapa. Masker N95/respirator partikulatb. Sarung tanganc. Pelindung wajah atau mata/face shieldd. Baju/coat/gaun/aprone. Tutup kepalaf. Sepatu tertutup atau memakai cover shoes

• Higiene petugas

• Kebersihan ruangan: dekontaminasi permukaan dengan desinfektan

TERIMA KASIH