SARS and the Clinical Laboratory
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1 SARS and the Clinical Laboratory Susan M. Poutanen, MD, MPH, FRCPC Microbiologist, Toronto Medical Labs & Mount Sinai Hosp. Infect. Dis. Clinician, University Health Network & Mount Sinai Hosp. Assistant Professor, University of Toronto, Canada March 3, APHL ID Conference Emerging ID Emerging Responses
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4 Worldwide SARS Outbreak eiris et al. Nature Medicine 004;10(12):S88-S97
5 Overview Overview of SARS Clinical Prediction Rules Diagnostics Tests Putting SARS into Context
6 Overview of SARS
7 SARS-associated Coronavirus (SARS-CoV) Enveloped Non-segmented, single-stranded, positive sense, RNA virus Kuiken et al. Lancet 2003; 362:
8 tadler et al. Nature Reviews Microbiology 2003; 1:
9 tadler et al. Nature Reviews Microbiology 2003; 1:
10 tadler et al. Nature Reviews Microbiology 2003; 1:
11 Peiris et al. Nature Medicine 2004;10(12): S88-S97
12 an der Hoek et al. Nature Medicine 2004;10(4):
13 oo et al. Journal of Virology 2005;79(2):
14 SARS Severe Acute Respiratory Syndrome acute onset respiratory syndrome typically moderate to severe asymptomatic and mild infection have also been detected
15 igure created by R. Chow for Poutanen and Low. Chapter 36. SARS n: Respiratory Infections (Eds Torres, Ewig, Mandell, Woodhead) Schematic of Symptom Progression fever/myalgia cough / dyspnea diarrhea 0 m 1 m 2 m
16 igure created by R. Chow for Poutanen and Low. Chapter 36. SARS n: Respiratory Infections (Eds Torres, Ewig, Mandell, Woodhead) Schematic of Symptom Progression fever/myalgia cough / dyspnea diarrhea 0 m 1 m 2 m
17 Clinical Outcome 20% admitted to ICU 15% required mechanical ventilation ~10% died Increased risk of death or ICU admission if: * Increased age * Co-morbidity * High LDH * High neutrophil count Tsui et al. EID 2003; 9: Fowler et al. JAMA 2003; 290: Lew et al. JAMA 2003; 290: Chan et al. Thorax 2003;58: Choi et al. Ann Int Med 2003;139:715-72
18 Clinical Prediction Rules
19 Clinical Prediction Rules Characteristics that increased the likelihood of SARS: Previous contact with a patient with SARS Fever, myalgia, malaise AbN CXR, abn lymphocyte and low platelet counts Characteristics that decreased the likelihood of SARS: Age 65 years or < 18 years Productive sputum Sore throat, rhinorrhea Abdominal pain High neutrophil count Leung et al. Ann Intern Med. 2004;141:
20 Clinical Prediction Rules Score translates to low or high risk group Hong Kong cohort Sensitivity 92% Specificity 63% Taiwan cohort Sensitivity 99% Specificity 52% Leung et al. Ann Intern Med. 2004;141: Ma et al. Ann Intern Med. 2005; 142:225-6
21 Diagnostic Tests
22 RT-PCR Positivity: plasma/serum NPA stool Symptoms: fever/myalgia cough / dyspnea diarrhea Transmissibility: IgG IgM Sero-positivity: 0 m 1 m 2 m Figure created by R. Chow fo Poutanen and Low. Chapter 36. SARS In: Respiratory Infections (Eds Torres, Ewig, Mandell, Woodhead)
23 0 m 1 m 2 m RT-PCR Positivity: plasma/sera NPA stool Viral Detection Symptoms: fever/myalgia cough / dyspnea diarrhea Transmissibility: IgG IgM Sero-positivity: Serologic Tests
24 Viral Detection
25 Viral Load: Peiris et al. Lancet 2003; 361:
26 han et al. EID 2004;10(2): RT-PCR Positivity:
27 rant et al. NEJM 2003;349:2468 RT-PCR Positivity of Plasma:
28 rant et al. NEJM 2003;349:2468 RT-PCR Positivity of Plasma: 79% positivity
29 igure created by R. Chow for Poutanen and Low. Chapter 36. SARS n: Respiratory Infections (Eds Torres, Ewig, Mandell, Woodhead) Schematic of RT-PCR Positivity Plasma / Serum NPA stool 0 m 1 m 2 m
30 igure created by R. Chow for Poutanen and Low. Chapter 36. SARS n: Respiratory Infections (Eds Torres, Ewig, Mandell, Woodhead) Schematic of RT-PCR Positivity Plasma / Serum NPA stool 0 m 1wk 2wk 3wk 1 m 2 m
31 igure created by R. Chow for Poutanen and Low. Chapter 36. SARS n: Respiratory Infections (Eds Torres, Ewig, Mandell, Woodhead) Schematic of RT-PCR Positivity Plasma / Serum NPA stool 0 m 1wk 2wk 3wk 1 m 2 m
32 igure created by R. Chow for Poutanen and Low. Chapter 36. SARS n: Respiratory Infections (Eds Torres, Ewig, Mandell, Woodhead) Schematic of RT-PCR Positivity Plasma / Serum NPA stool 0 m 1wk 2wk 3wk 1 m 2 m
33 OC43/229E CoV Nasal Wash RT-PCR Viral Load an Elden et al. JID 2004; 189(4):652-7
34 OC43/229E CoV Nasal Wash RT-PCR Viral Load an Elden et al. JID 2004; 189(4):652-7
35 Viral Detection - Specimens Week 1: plasma/serum, respiratory specimens* Week 2: respiratory specimens*, stool Week 3: stool Multiple specimens over time * LRT higher yield compared to URT samples
36 Respiratory Specimens heng et al. Lancet 2004;363:
37 Viral Detection Post-Mortem Samples Lung Bowel Lymph node Spleen Liver
38 Viral Detection Post-Mortem arcas et al. JID 2005;191:
39 Viral Detection - Methods Culture Nucleic Acid Amplification Testing (NAAT) Other (e.g. for tissue analysis) Electron microscopy, immunohistocytochemistry, in situ hybridization
40 Culture Culture Requires level 3 laboratory Not as sensitive as NAAT more readily isolated from resp tract than stool; most successful during the first 2 weeks of illness han et al. EID 2004;10(2):
41 Nucleic Acid Amplification Testing RT-PCR Multiple targets polymerase, nucleocapsid, spike genes Variations Traditional vs. real-time platform In-house vs. commercial Microarray-based Bead-based Extraction methodology
42 rosten et al. EID 2004;10(12): WHO EQA Study Factor # Labs P Value
43 Viral Detection - WHO HO. SARS Guidelines. October 2004
44 Viral Detection - CDC DC. SARS Laboratory Guidance. May 2004
45 Serologic Tests
46 igure created by R. Chow for Poutanen and Low. Chapter 36. SARS n: Respiratory Infections (Eds Torres, Ewig, Mandell, Woodhead) Schematic of Sero-Positivity IgG IgM 0 m 1 m 2 m
47 SARS-CoV IgG Kinetics Cumulative % of patients with eroconversion D28 Peiris et al. Lancet 2003; 361: Time after onset of symptoms
48 Chen et al. JID 004;189: SARS-CoV IgM and IgG Kinetics
49 SARS-CoV IgM and IgG Kinetics IgG IgM Shi et al. J Clin Virology 3004;31: 66-68
50 Serologic Testing - Specimens Acute serum Convalescent serum defined as >28 days post symptom onset
51 Serologic Testing - Methods Immunofluorescent assays Enzyme immunoassays Western Blot Immunodot Assay Neutralization assay
52 Serologic Testing - WHO HO. SARS Guidelines. October 2004
53 Serologic Testing - CDC DC. SARS Laboratory Guidance. May 2004
54 Other Diagnostic Tests
55 Antigen Capture EIA Antigen capture EIA based on monoclonal antibodies against nucleocapsid protein Sensitivity 94% using serum first 5 days Specificity 99.9% he et al EID 2004;10(11):
56 he et al EID 2004;10(11):
57 SELDI-TOF Surface-enhanced laser desorption/ionization time-of-flight Diagnostic patterns Prognostic patterns ang et al and Yip et al Clin Chem 2005; 51: and 56-64
58 ang et al Clin Chem 2005; 51: 47-55
59 ip et al Clin Chem 2005; 51: 56-64
60 Putting SARS into Context
61 Will SARS Return? How SARS might return: Humans Animals Research Laboratories
62 CDC DC. SARS Laboratory Guidance. Jan 2004
63 CDC Testing should be performed only at high risk for SARS-CoV disease DC. SARS Laboratory Guidance. Jan 2004
64 Rule out Testing Cannot be used to rule out SARS Co-infections with hmpv, influenza virus May help identify cause of clusters han et al. EID 2003;9; chrag et al. EID 2004;10(2);
65 Rule out Testing anadian Public Health Laboratory Network Nov 2003
66 Add-ons hmpv?non-sars coronaviruses?rhinoviruses?others
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