Laboratory Diagnostics Bonna Cunningham, MS North Dakota Public Health Laboratory.
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Transcript of Laboratory Diagnostics Bonna Cunningham, MS North Dakota Public Health Laboratory.
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Laboratory Diagnostics
Bonna Cunningham, MSNorth Dakota Public Health Laboratory
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SARS Testing* at CDC
Antibody TestsMolecular TestCell Culture
* SARS serologic and molecular assays will be available at the NDPHL shortly (pending release by CDC).
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Antibody Tests
IFA and ELISA Reliable 21 days post onset of
feverAntibodies detected as early as
14 days in some cases
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Molecular Test
RT-PCR Positives reportedNegatives repeated with more
sensitive primers when available
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Viral Culture
Respiratory secretions and bloodVero, Vero E6 support virus
replicationOther cell lines being evaluated
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Interpreting Test Results
• Positive Indicates current or recent infection with the coronavirus.
•Negative Does not mean the patient does not have SARS. Diagnose on clinical evaluation and possible past exposure.
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Potential SARS SpecimensSerum 5-10 ml blood in serum separator
EDTA whole blood 5-10 ml
Stool 10-50 cc
NP swabs/OP swabs Dacron swab in viral transport
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M4 Viral Transports
Insert illustration
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Location of M4 Viral Transports in North Dakota
Two M4 viral transports/swabs in each smallpox shipper Four shippers at each NDLRN Level A
laboratory Four shippers at each District Health Unit
Additional six M4 viral transports/swabs at each District Health Unit
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Level A Labs
Insert MAP
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District Public Health Units
Insert MAP
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Packaging and Shipping
Follow IATA/DOT packaging regulations for Diagnostic Specimens*
http://www.cdc.gov/ncidod/sars/packingspecimens-sars.htm
* “Smallpox shippers” issued by NDDoH meet requirements
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Smallpox Shippers
(Insert Illustration)
•Contact the NDPHL for assistance–Phone Number: 701.328.5262
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Laboratory BiosafetyEstablish protocols to protect laboratory workers Labeling suspected SARS cases Handling blood specimens for routine testing Handling specimens for microbiological
analysis Define BSL-2 practices* Define BSL-3 practices*
*Refer to CDC/NIH Biosafety in Microbiological and Biomedical Laboratories manual (BMBL):
http://www.cdc.gov/od/ohs/biosfty/bmb14/bmb143s3.htm
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Blood Specimens for Routine Testing
Use universal precautionsWear appropriate PPEDisposable glovesLab coatEye/face shields
Use safe centrifugation practices
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Centrifuging ProtocolsUse sealed centrifuge cups or rotors Load and unload in BSC
If sealed centrifuge cups and BSC not available Keep testing to a minimum Centrifuge separately Limit number of staff in room where
centrifuge is located Use respiratory protection when unloading
centrifugeN-95 maskEye/face shields
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BSL-2 Activities
Exam/processing of formalin-fixed tissuesMolecular analysis of extracted prepsEM with glutaraldehyde-fixed gridsRoutine exam of bacterial/mycotic culturesRoutine staining/analysis of fixed smearsPackaging specimens for transport
--Interim Laboratory Biosafety Guidelines for Handling and Processing Specimens Associated with SARS, Department of Health and Human Services, Centers for Disease Control and Prevention, April 2, 2003
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BSL-2 Activities/BSL-3 Practices
Aliquoting/diluting specimensInoculating bacterial/mycotic culture mediaMicrobiology testing other than propagation of viral agentsNucleic acid extractions of untreated specimensPrep/fixing of smears for micro analysis
--Interim Laboratory Biosafety Guidelines for Handling and Processing Specimens Associated with SARS, Department of Health and Human Services, Centers for Disease Control and Prevention, April 2, 2003
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BSL-3 Activities
Viral cell culture
Initial characterization of viral agents in cultures of SARS specimens
--Interim Laboratory Biosafety Guidelines for Handling and Processing Specimens Associated with SARS, Department of Health and Human Services, Centers for Disease Control and Prevention, April 2, 2003
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NDDoH Website
http://www.health.state.nd.us/disease/SARS
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Burleigh
Oliver
Dunn
Slope
Bowman
BillingsGolden Valley
Stark
Hettinger
Adams Sioux
Grant
Mercer
Morton
Mountrail
Williams
McKenzie
Divide Burke
McHenry
McLean
Ward
RenvilleBottineau
Kidder
Dickey
Emmons
McIntosh
Stutsman
LoganLa Moure
Sargent
Richland
Barnes
Ransom
Cass
Ramsey
Eddy
WellsSheridan
Foster
Rolette
Pierce
Benson
Towner
Nelson
Steele
Griggs
Traill
Grand Forks
Cavalier
Walsh
Pembina
North Dakota Laboratory Response Network
12
8
9 9
7
6
51 2
13141516
Level-A Laboratories
1. Mercy Hospital, Williston
2. Trinity Med. Cen., Minot
3. USAFB, Minot
4. Presentation Hospital, Rolla
5. Mercy Hosp, Devils Lake
6. USAFB, Grand Forks
7. Altru Hospital, Grand Forks
8. Innovis Health Center, Fargo
9. MeritCare Med Cen., Fargo
10. VA Medical Center, Fargo
11. Dakota Clinic, Fargo
12. Mercy Hosp., Valley City
13. Health Care Hosp., Jamestown
14. MedCenter One, Bismarck
15. St. Alexius Med. Cen., Bismarck
16. St. Joseph Hospital, Dickinson
17. West River Reg. Med. Center, HettingerNDPHL
3 4
17
10
11
Level-B/C Laboratory
North Dakota Public Health Laboratory (NDPHL)
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Larry A. Shireley, MS, MPHState Epidemiologist
North Dakota Department of Health
Severe Acute Respiratory Severe Acute Respiratory Syndrome (SARS)Syndrome (SARS)
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Onset since February 1, 2003 Measured temperature ≥ 100.50F Respiratory Illness*
AND Travel within 10 days of symptoms onset to:
Peoples’ Republic of China, Hong Kong, Hanoi, Viet Nam or Singapore OR
Close contact within 10 days of symptoms onset to: Suspected SARS case Respiratory illness & travel to above areas
* WHO definition requires radiographic evidence of infiltrates consistent with pneumonia or respiratory distress syndrome
CDC SARS Case DefinitionCDC SARS Case DefinitionApril 10, 2003April 10, 2003
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Epidemiology
Transmission Person – Person Health Care Workers Community Transmission
United States – Primarily related to travel
Primarily adults – 25 – 70 Uncommon < 15 years old
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Epidemiology
Most Cases Resolve ~90%+ day 6-7 Mortality ~ 4 %
United States Cases less severe Reasons?
Cultural? Medical care? Other co-infection?
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SARS Time Line
November 16, 2002 Index Case – Guangdong, China (Reported Feb 14, 2003)
Feb 11, 2003 - First Case Reports from China Feb 21 Hong Kong hotel outbreak Feb 28, 2003 – Viet Nam reports cases Global Alert – March 12, 2003 March 14 – Canada reports cases March 15 – WHO Travel Advisory March 24 – Link to coronavirus April 3 – CDC Travel Advisory April 4 – Executive Order - Quarantine
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Suspected Cases Worldwide(April 12, 2003)
Total Cases 2,960 Deaths 119
Number of Countries 19
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Suspected SARS Cases by Country
April 12, 2003
Country Cases
New Cases Deaths Recovered
Local Transmission
Brazil 2 0 0 0 No
Canada 101 3 10 26 Yes
China 1,309 19 58 1,037 Yes
Hong Kong
1,108 49 35 215 Yes
Taiwan 23 2 0 7 Yes
France 5 0 0 1 No
Germany 6 0 0 4 No
Ireland 1 0 0 1 No
Italy 3 0 0 2 No
Japan 4 0 0 0 No
Kuwait 1 0 0 0 No
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Country Cases
New Case
s
Deaths
Recovered
Local Transmissio
n
Malaysia 4 0 1 0 No
Romania 1 0 0 NA No
Singapore 147 14 9 77 Yes
South Africa 1 0 0 0 No
Spain 1 0 0 0 No
Switzerland 1 0 0 1 No
Thailand 8 1 2 5 No
United Kingdom
6 1 0 3 Yes
United States 166 0 0 NA Yes
Viet Nam 62 1 4 46 Yes
Total 2,960 90 119 1,425
Suspected SARS Cases by Suspected SARS Cases by CountryCountry
April 12, 2003 (cont)April 12, 2003 (cont)
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5 RI 1
CT 4
NJ 3
MA 5
35
21
5
1
5
12
7
13
1 1
6
3
2
3
HI 5
5NH 1
6
14
VT 2
5
11
1
1
Reported Suspect Cases of SARSReported Suspect Cases of SARS United States United States
through April 10, 2003through April 10, 2003
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Characteristics of US SARS Cases*
As of April 9, 2003
135 (81%) Adults
154 (93%) Travel to endemic area 9 (5%) Household contact to
SARS 3 (2%) Health Care Workers 60 (36%) Hospitalized >24 hours 33 (20%) Radiographic
abnormalities*166 cases
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Number of Suspected Cases of SARS by Exposure Category and Date of Illness
Onset United States, 2002
CDC, MMWR April 11, 2003
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Number and Percentage of Reported SARS Number and Percentage of Reported SARS Cases by Selected Characteristics Cases by Selected Characteristics
United States, 2003United States, 2003
* N = 166.
†To mainland China, Hong Kong, Hanoi,
or Singapore.
§ As of April 9, no deaths of SARS patients
have been reported in the United States.
¶ Respiratory distress syndrome.
CDC, MMWR April 11, 2003
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Keys to Control
Early recognition and treatment of cases Stringent Infection Control Procedures in Hospitals and Clinics Prompt Reporting of Suspected Cases Investigation & Contact Tracing Public Awareness and Education
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SARSSevere Acute Respiratory Syndrome
Clinical IssuesClinical Issues
“We've never faced anything on this scale with such a global reach.”
-Dr. David Heymann, World Health Organization
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SARS Background
26 Feb 03 1st case HanoiWHO official - Dr. Carlo Urbani
died 29 Mar 03 SARS
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SARS BackgroundCase 1
Disease symptom onset Feb. 15
Traveled from Guangdong Province to Hong Kong Hotel M Feb 21
Died Feb 23
4 health care workers and 2 family contacts, and 10 hotel guests developed disease
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SARS Background Case 2
Admitted to a Hanoi hospital Feb 26 Travel to Hong Kong Hotel MRespiratory failure requiring ventilatory supportEvacuated to Hong Kong; died March 1259 contacts developed disease
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MMWR March 28, 2003 / 52(12);241-248
Chain of Transmission at Hotel M - Hong Kong 2003
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MMWR April 4, 2003 / 52(13);269-272
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MMWR March 28, 2003 / 52(12);241-248
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Hong Kongstudy of 50 cases
www.thelancet.com 8 Apr 03
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Predictors of “severe” SARS in Hong Kong www.thelancet.com 8 Apr 03
Complicated(n=19)
Uncomplicated(n=31)
p
age 49.5 39 0.005
comorbidity 5 1 0.05
DM
Chronic active hepatitis
Cardiomopathy
HTN
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Predictors of “severe” SARSwww.thelancet.com 8 Apr 03
Method of contactTravel to chinaHCWHospital visitHousehold contactSocial contact
P = 0.09
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Predictors of “severe” SARSwww.thelancet.com 8 Apr 03
Duration of symptoms before admission ~ 5 daysTemperature on admission 38.8WBCInitial lymphocyte 0.66 vs .85ThrombocytopeniaImpaired LFT’s 11 vs. 6
P=0.04
P = 0.01
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Predictors of “severe” SARSwww.thelancet.com 8 Apr 03
Complicated Uncomplicated p
# pt on ribivirin andsteroids (R&S)
18 31
Mean days to startR&S
7.7 5.7 0.03
Start R&S afterworsening
12 0 0.0001
Response to R&S 11 28 0.02
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Hong Kong
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Hong Kong
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DemographyHong Kong
Total 138Female 72HCW 69Doctors 20Nurses 34Allied health workers 15Medical Students 16Patients 19Relatives 34
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Common SymptomsHong Kong
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Hong Kong
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Serum ChemistryHong Kong
Elevated LDH 71%Elevated CPK 32%median 126 U/L, range: 29-4644
Elevated ALT 23%
Hypokalemia 25%Hyponatremia 20%
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Hong Kong
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Hong Kong
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It is important to to consider other pathogens: influenza etc
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CXR ResolutionHong Kong
In 7 days median duration:
82% of patients had 25% resolution of chest shadows
69% of patients had 50% resolution of chest shadows
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Lessons Learned
Early high dose steroid is worthyRibivirin may be beneficialDon’t use nebulizerDon’t use non-invasive positive pressure ventilationChest physiotherapy may help
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Identification of Severe Acute RespiratorySyndrome in Canada
published at www.nejm.orgon March 31, 2003
ER, 2 meters away
Diabetic
Diabetic, died at home
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Severe Acute Respiratory Syndrome in Canada
published at www.nejm.org on March 31, 2003
No SARS contact
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Clinical Features of the Canadian Patientswith SARS at Presentation
published at www.nejm.orgon March 31, 2003
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Summary of the 20 cases published at www.nejm.org on March 31, 2003
Incubation period 1 to 11 daysmedian 5 days
Fever 100%Most patients:Rigor, nonproductive cough,
dyspnea, hypoxia, malaise, and headache
Lung crackles and dullness on percussion
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Summary of the 20 cases published at www.nejm.org on March 31, 2003
LymphopeniaElevated transaminasesHypoxiaCXR and CT scansSimilar to interstitial pneumoniaProgressive bilateral air space
disease
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Summary of the 20 cases published at www.nejm.org on March 31, 2003
Majority of cases suggest droplet transmissionIndex casesFamily membersHCW’s
failure to follow infection controls
Fourth - and fifth generation of casesWill blur epidemilogical links
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Summary of cases
Increase morbidity and mortalityadvance agecomorbidities e.g. DM
Ribivirin and prednisone early may be of benefit
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Recommended Protocol for Clinical Treatment
Community acquired pneumonia protocol1. R/O influenza2. Consider atypicals3. Ribaviran and Prednisone4. No aerosolized procedures
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Prognosis of SARS
~ 3 - 4% mortality
6% survive but prolong, complicated course
90% recover
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CoronavirusEtiology of SARS ?
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Coronavirus in culture
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Serological Evidence of Coronavirus
Found in multiple geographic areasHong Kong - 9 ptsUSA - 1Bangkok - 1Singapore - 4
Seropositivity occurs ~ 11 to 24 days after onset
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Multiple Methods Point to Coronavirus
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Genetic Evidence for Coronavirus
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CoronavirusEtiology of SARS ?
Increase confidence in CoronavirusNew case definition anticipated
To include laboratory test criteria
International testing of antiviral compounds
Vaccine research underway
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SARS Unresolved Issues
? Airborne transmissionextensive spread within buildings in
Asia
Fomite transmissionCoronavirus can survive in the
environment for a few hoursCoronavirus found in animal stools
No proven, successful population based strategy prevention
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SARS Optimism for future control
Effective coronavirus vaccines in animalsNovel antiviral drugs may be found
Infection control measures work
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Infection Control
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Administrative
CommunicationEducatePolicies & proceduresEnforcement
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Personal Protective Measures
MaskGloves and gowns Eye protectionHand hygiene
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PrinciplesHypertransmitters - some patientsProtection of patients, staff, visitorsPrevent spread in the facility and communityTarget all modes of transmission until SARS epidemiology is understoodProtect facilities so routine care is not impaired
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Triage for SARS in Ambulatory Care
Targeted screeningCurrently:Travel historyContact with a person with SARSAir travel to a country with SARSFever and or respiratory
symptoms
www.cdc.gov/ncidod/sars/triage_interim_guidance.pdf
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Triage for SARS in Ambulatory Care
Evaluate in a separate assessment areaIf SARS suspected: Patient wears a surgical mask HCW applies Airborne and Contact
PrecautionsN95 if available; at least a surgical maskGlovesGownEye protectionNegative pressure if available
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Respiratory Protection
Patient Cover coughs
with tissue or hand
Surgical mask Hand hygiene
Healthcare Workers N95 PAPR
Surgical mask if respirator not available
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Engineering measures
Control of ventilationControl of trafficSecurity
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Aerosolizing Procedures for SARS
Evaluate patients for SARS before: Aerosolized medication treatmentsSputum inductionBronchoscopyAirway suctioningEndotracheal intubation
Perform only if medically necessaryUse Airborne Precautions as per TB
www.cdc.gov/ncidod/sars/pdf/aerosolinfectioncontrol-sars.pdf
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Visitor Restrictions
Symptomatic close contacts of SAR patients should not enter facility.
Screening.
Educate visitors about precautions if visiting a SARS patient.
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Post-mortem
Standard Precautions GownN95, N100, or PAPR (preferred for
aerosolizing procedures)AutopsyMinimum 12 ACH and negative
pressurePrevent percutaneous injuryDispose of PPE carefully
www.cdc.gov/ncidod/sars/pdf/sarsautopsy.pdf
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Patients with suspected SARS and Household Contacts
Limit interactions outside the home until 10 days after resolution of symptomsHand hygieneGlovesPatient covers coughs with tissue or maskDo not share utensils, towels, beddingClean surfaces with disinfectantHousehold contacts do not limit activity outside the home if asymptomatic
www.cdc.gov/ncidod/sars/pdf/ic-closecontacts-sars.pdf
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Exposure Management
Definitions Exposure:Travel from areas with documented or suspected community transmission of SARSClose Contacthaving cared forhaving lived withhaving direct contact with respiratory
secretions and/or body fluids
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www.cdc.gov/ncidod/sars/pdf/exposuremanagement-sars.pdf
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Exposure Management in Healthcare
Transmission associated with unprotected exposureExclude from duty if symptomatic within 10 days of exposure to SARS. Continue until 10 days after resolution of symptoms.Screen exposed daily for fever and respiratory symptoms.Facilities with SARS patients: educate workers about symptoms passive surveillance
www.cdc.gov/ncidod/sars/pdf/exposureguidance.pdf
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School Children Exposed to SARS
No symptoms-do not exclude from school but monitor symptomsFever or respiratory symptoms within 10 days of exposure Stay home; if no progression to SARS, then
return to school If progresses to SARS, precautions
continued until 10 days after resolution Alternative housing for students in dorms,
etc.
www.cdc.gov/ncidod/sars/pdf/exposurestudents.pdf
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Advice for Travelers
Know about SARS in the travel areaDo not go to China, Hong Kong, Singapore or Hanoi unless necessary.No advisories about Canada.Current immunizations.Hand hygiene; bring alcohol hand rubsSeek medical attention if ill
www.cdc.gov/ncidod/sars/pdf/travel_advice.pdf
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SARS Infection Control at Altru Phase 1
Identify and rapidly isolate initial patients Signs at entry: passive
screening First contacts screen
for travel and SARS exposure
EOD: active screening SARS Call Center Use existing negative
pressure rooms Education
S A R S A L E R TS e v e r e A c u t e R e s p i r a t o r y S y n d r o m e
P L E A S E P U TO N A M A S K I F :
Y o u h a v e t r a v e l e d t o a n y o f t h e s e a r e a si n t h e l a s t 3 w e e k s :
• A s i a , i n c l u d i n g :C h i n a , H o n g K o n g , H a n o i , V i e t n a m , o r S i n g a p o r e
• T o r o n t o , C a n a d a
A N D
Y o u a r e i l l w i t h :
F E V E R h i g h e r t h a n 1 0 0 . 4 ° F R e s p i r a t o r y i l l n e s s
- C O U G H- S h o r t n e s s o f b r e a t h- D i f f i c u l t y b r e a t h i n g- R e s p i r a t o r y d i s t r e s s
O R
H a v e h a d c l o s e c o n t a c t w i t h a p e r s o n k n o w n o rs u s p e c t e d t o h a v e S A R S .
A P R I L 7 . 2 0 0 3
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Summary
Use epidemiologyPassive and active screening Use standard, airborne, and contact precautions