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Pengurusan Sample MERS-COV


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Taklimat berkenaan MERS-COV (Middle East Respiratory Syndrome-Corona Virus) berkenaan pengurusan sample yang diambil dari pesakit.

Virus ini mula tersebar di Arab Saudi, dan perhatian lebih perlu diberikan kepada jemaah-jemaah yang baru pulang dari Umrah di Makkah & Madinah, Arab Saudi.

Published in: Health & Medicine, Technology
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Pengurusan Sample MERS-COV

  1. 1. Pengurusan Sample Bagi Kes Jangkitan “Middle East Respiratory Syndrome- Coronavirus” (Mers- CoV) 1 Ref:NIAID
  2. 2. • Background of the Virus • Samples suitable for testing and WHO Guidelines • Real time RT-PCR for Mers-CoV • Main Hospitals and MKAK Sg Buloh and MKA Kota Kinabalu Overview
  3. 3. Virus classification -RNA viruses
  4. 4. Middle East Respiratory Syndrome- Coronavirus (Mers-CoV)  First reported in June 2012, Saudi Arabia  Coronaviridae Family, Genus Betacoronavirus (same as SARS-CoV).  Positive sense, single stranded RNA virus  About 26-32 Kb with “crown-like” feature  Transmission- Close Contact -aerosol (droplet), further investigation. (SARS-CoV- “unique pathogenesis” as involved both upper and lower respiratory tract and gastro enteritis)
  5. 5. WHO Guidelines (27 June 2013)  Generally incubation period for cases of strong suspicion is less than 1 week  BUT in at least 1 case, known exposure occurred 9 to 12 days prior to onset of illness Longer period of observation for contacts of cases  Incubation period may exceed 1 week but is less than 2 weeks
  6. 6. Source of Clinical Specimens Hospitals – Inpatients with severe Influenza NO clinical specimens from outpatients and contacts!
  7. 7. Preferred Sample For Testing (WHO Guidelines, 27 June 2013)  Lower Respiratory Tract  Sputum  Endotracheal Aspirate  Bronchoalveolar Lavage  If pt without signs and symptoms of lower respiratory tract infection and if lower respiratory tract specimen NOT Possible or Clinically Indicated Both Nasopharyngeal and Oropharyngeal specimens should be collected
  8. 8.  If initial testing of Nasopharyngeal swab is NEGATIVE BUT pt is Strongly Suspected of Mers-CoV, pt MUST be RE-TESTED using Lower Respiratory Tract Specimen  Others: tissue from biopsy/autopsy including lung, whole blood (in EDTA), paired serum for serology (4 weeks apart)
  9. 9. When to collect specimen • As soon as possible after symptoms begin (24-72hrs) • Best before antiviral medications are administered
  10. 10. Swabs • Ideal – synthetic tip • Specimens collected with swabs made of calcium alginate, cotton or wooden sticks are NOT ACCEPTABLE (inhibit PCR)
  11. 11. Sample collection Nasopharyngeal swab
  12. 12. Transport media - Plain sterile bottles for aspirates and sputum - VTM for swabs - Plain tubes for paired sera
  13. 13. Viral Transport Media (VTM) • Place swabs in sterile collection vials containing 1-2 ml of VTM • Vials can be stored in a freezer at -20 o C until use • Vials can be stored for short periods of time at 4 – 6o C
  14. 14. Storage and transport of specimens • Transport to the laboratory as soon as possible • Store specimens and transport at 4o C • Store specimens at -70o C beyond 48 hours. • If a -70°C freezer is not available, specimens should be kept at 4°C, preferably no longer than 4 days. • Freezing at higher temperatures (e.g., -20°C) can reduce the likelihood of virus detection.
  15. 15. 3 layer packaging
  16. 16. 1 2 3
  17. 17. • Shipping clinical specimens: - Clinical specimens should be shipped in ice (cold chain) in appropriate packaging - All specimens should be labeled clearly and include information requested .
  18. 18. Transportation of Samples to Testing Facilities Clinicians/Health care providers advised to wear appropriate PPE during sample taking from patients All secretions should be placed in sterile bottle and t transported on ice All swabs must be placed in sterile tubes with sufficient VTM and transported on ice. All samples must use “triple packaging” format with request forms before being transported to testing facilities
  19. 19. Issues
  20. 20. Laboratory Testing Methods 1. Real time RT-PCR (molecular lab) 2. Cell Culture 3. RT-PCR & Sequencing 4. Serology (paired sera 4 weeks apart)
  21. 21. Lower Respiratory Tract (Sputum, Endotracheal Aspirate, Bronchoalveolar Lavage) Other respiratory tract samples (Both Nasopharyngeal and oropharyngeal specimens), T/S IMR will provide laboratory testing for these facilities until 01/08/2013
  22. 22. False Negative Results: Samples NOT from Lower Respiratory Tract (request for fresh sample from lower respiratory tract if HIGH disease suspicion index) Poor quality of specimen/ PCR inhibition  Specimen was collected late or very early in the illness  The specimen was not handled or shipped appropriately
  23. 23. Quality Control of Testing during rRT-PCR  In the event of “positive results”, Samples should be referred to IMR for confirmation (sequencing)- only in INITIAL stage  IMR will provide Quality Assurance Programme for all testing facilities this year onwards for rRT-PCR Mers-CoV.
  24. 24. Briefing Testing Procedures
  25. 25. List Of Facilities Trained To Perform rRT-PCR for Mers-CoV 1. Makmal Kesihatan Awam Kebangsaan, Sg Buloh 2. Makmal Kesihatan Awam Kota Kinabalu 3. Hospital Sungai Buloh 4. Hospital Kuala Lumpur 5. Hospital Umum Sarawak, Kuching 6. Hospital Pulau Pinang 7. Hospital Sultanah Aminah Johor Bahru 8. Hospital Raja Permaisuri Bainun Ipoh 9. Hospital HTAA, Kuantan 10.Hospital Melaka 11.Hospital HSNZ Kuala Trengganu 12.Hospital HRPZ II Kota Bharu 13.Hospital Sultanah Bahiyah Alor Setar 14.Hospital Tuanku Jaafar Seremban Tentative Start Date for Testing : 01/08/2013