PulseNet Gestalt. John Besser Minnesota Department of Health

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1 PulseNet Gestalt John Besser Minnesota Department of Health

2 Questions I Was sked to ddress How are epidemiologic investigations affected by 2 nd enzyme data? re investigations delayed until results are available? Does delay affect the outcome of investigations? re resources wasted if 2 nd enzymes later reveal that multiple patterns were found and therefore a common source is less likely? How often are epi clusters found that do not correlate with PFGE 2 enzyme clusters? re further investigations conducted or additional lab testing done (3 rd enzyme, MLV for salmonella)? What happens if no epi links can be found for 2 enzyme clusters? What effect does the inclusion of unrelated cases have on epi analyses such as hypothesis generation, odds ratio, etc?

3 Implications of the Two Enzyme Policy Sensitivity, specificity, and significance issues Public health/regulatory action Practical implications

4 PulseNet Gestalt ge stalt or Ge stalt ( P ) Pronunciation Key (gshtält, -shtôlt, -stält, -stôlt) n. pl. ge stalts or ge stalt en (-shtältn, -shtôltn, -stältn, -stôltn) physical, biological, psychological, or symbolic configuration or pattern of elements so unified as a whole that its properties cannot be derived from a simple summation of its parts.

5 Specificity Sensitivity discrimination The use of 2 enzymes increases subtype specificity

6 Direct Contact with Primary Reservoir

7 Transmission via Vehicle

8 Ingredient in Product amplification

9 Person-To-Person Transmission

10 Secondary Transmission

11 Person-To-Pet Transmission

12 Multiple Transmission Routes

13 Multiple Transmission Routes Environment Patron illness Contaminated product Workers Other foods

14 Multiple Transmission Routes 12 Number of Cases J FMMJJ SONDJ FMMJ JSOND J FMMJ J SOND JFMMJ JSOND

15 PFGE Case Definition

16 Goal: Identification of transmission event Method: marker for short-term evolutionary relationships minutes

17 conjugation transduction transformation mutation Via transposons plasmids integrons phage transcription errors

18

19 Subtyping (for hypothesis generating and testing): Refines the case definition to increase signal:noise

20 Refining the Case Definition One enzyme PFGE match Outbreakassociated

21 Refining the Case Definition Tenover criteria Outbreakassociated

22 Refining the Case Definition One enzyme PFGE match Outbreakassociated

23 Refining the Case Definition Two enzyme PFGE match Outbreakassociated

24 Refining the Case Definition Ten enzyme match Outbreakassociated

25 inflection point Type II error Type II error significance specificity of case definition

26 Specificity Sensitivity discrimination (Example: Bacteria not identified at all) ll inclusive ll isolates identical

27 Specificity Sensitivity (Example: Every isolate completely sequenced) ll exclusive ll isolates different discrimination

28 Gram Rxn genus species Tenover group subtype subtype, 2 enz Specificity Sensitivity discrimination

29

30 inflection point Type II error Type II error significance specificity of case definition

31 Gram Rxn genus species Tenover group subtype subtype, 2 enz Specificity Sensitivity discrimination How much discrimination is the right amount of discrimination?

32 It Depends On. The question you re trying to answer Prevalence of exposure Prevalence / type of agent Number of cases Other factors which contribute to specificity of case definition

33 Question: What is the vehicle for illnesses in a recognized outbreak?

34 Question: re two recognized outbreaks connected?

35 Question: What is the extent of an outbreak?

36 Question: Is there a clonal relationship between strains in different locations? Case Definition: Y. pestis isolated (biovar orientalis) from different geographic areas Burma India Indonesia US Vietnam Courtesy Kristy Kubota, CDC; ICEID Poster k2006

37 Question: Is their a common source for a cluster of cases?

38 For hypothesis generating and testing: In general, more subtype specificity is better..to a point

39 Factors That ffect the Need for Specificity Prevalence of exposure Prevalence / type of agent Number of cases

40 Odds Ratio exp+ exp- case C control B D OR = D CB

41 Prevalence of Exposure exp+ exp- case C control B D OR = D C B Odds ratio: inversely proportional to odds ratio

42 Low Prevalence Exposure case control exp+ exp- C B D Example: my mother s fruit cocktail surprise OR = D CB Consequence: less cases needed to reach significance

43 High Prevalence Exposure case control exp+ B Example: tomato consumption exp- C D OR = D CB Consequence: more cases needed to reach significance

44 Specific Exposure Definition case control exp+ exp- C B D Example: tomato consumption, supplier X OR = D CB Consequence: less cases needed to reach significance

45 Specificity and ssociation Exposure: tomatoes Case: S. Montevideo Control: not ill Exposed Not Exposed controls cases

46 Specificity and ssociation Exposure: tomatoes Case: S. Montevideo Control: not ill Case Control Exposed 6 3 Not Exposed 5 4 controls odds ratio = 1.6 cases

47 Specificity and ssociation Exposure: source tomatoes Case: S. Montevideo Control: not ill Tomato source Tomato source B Store X Store Y controls cases

48 Specificity and ssociation Exposure: source tomatoes Case: S. Montevideo Control: not ill Case Control Exposed 5 1 Not Exposed 6 6 controls odds ratio = 5 cases

49 Refining the EXPOSURE information: Increases signal noise ratio

50 exp+ case control B exp- C D OR = D CB

51 Case Classification exp+ Outbreak case control B exp- C C D Non-outbreak exp+ exp- C

52 Exposure Ratio Outbreak case control exp+ B exp- C C D Non-outbreak Ratio = specificity of case definition exp+ exp- C ffected by.. Prevalence of agent Specificity of agent definition Other case definitions

53 exp+ Outbreak Exposure Ratio case control B exp- C C D Non-outbreak exp+ exp- C

54 Low Prevalence gent case control exp+ exp- C B D Example: Salmonella Poona OR = D CB Few misclassified cases

55 High Prevalence gent case control exp+ exp- C B D Example: S. Typhimurium common subtype OR = D CB Many misclassified cases

56 More Specific Case Definition: 2 Enzymes case control exp+ exp- C B D Example: S. Typhimurium Subtype X1, B7 OR = D CB Less misclassified cases

57 Specificity and ssociation Exposure: tomatoes Case: S. Montevideo Control: not ill Exposed Not Exposed controls cases

58 Specificity and ssociation Exposure: tomatoes Case: S. Montevideo subt. Control: not ill Exposed Not Exposed controls cases

59 Specificity and ssociation Exposure: tomatoes Case: S. Montevideo subt. Control: not ill Case Control Exposed 5 3 Not Exposed 1 4 controls odds ratio = 6.7 cases

60 Specificity and ssociation Exposure: source tomatoes Case: S. Montevideo subt. Control: not ill Case Control Exposed 5 3 Not Exposed 1 4 controls odds ratio = 6.7 cases

61 Specificity and ssociation Exposure: source tomatoes Case: S. Montevideo subt. Control: not ill Case Control Exposed 5 1 Not Exposed 1 6 controls odds ratio = 30 cases

62 Specificity and ssociation Case Definition Non-specific Case Definition Specific Exposure Non-specific OR=1.6 OR=6.7 Exposure Specific OR=5 OR=30

63 Timeliness of the interview exp+ exp- case C control B D OR = D CB affects the sensitivity ND specificity of exposure information

64 E. coli O157:H7 Subtype Surveillance; Minnesota, No. of cases Jan Feb Mar pr May Jun Jul ug Sep Oct Nov Dec Month

65 Statistical ssociation with Consumption of Hamburger from Producer ; November- December No. of cases No. of cases Nov Dec Nov Dec Without PFGE OR = 1.93; p = NOT significant- With PFGE OR = 17.1; p = SIGNIFICNT-

66 Implications of the Two Enzyme Policy Sensitivity, specificity, and significance issues Public health/regulatory action Practical implications

67 E. coli O157:H7 Outbreak ssociated with Pre-packaged Salad: Whole Different Timeline MDH issues Press Release 3:29 P.M. from Seattle law firm to MDH 3:56 p.m. Contact person for outbreak posted on law firm website 4:45 p.m. Minneapolis Law Firm Star Tribune d Solicitation First lawsuit filed in Minneapolis Second lawsuit filed in Minneapolis September October 2005

68

69

70 Microbial DN Fingerprinting : dependent on continuous genetic change

71 Question: Does the DN in a specimen match the DN in an individual? Human DN testing X Year

72 Crime investigation: 22 Odds of exposure given illness Outbreak investigation: No. of cases in With PFGE OR = 17.1; p = 0.005

73 Multi-State Salmonella Typhimurium Outbreak ssociated with Cold Stone Creamery Ice Cream gent: rare (never seen before subtype), multiple cases, multiple states Temporal relationship: 10 cases/1 week Exposure: rare ( cake batter ice cream )

74 Implications of the Two Enzyme Policy Sensitivity, specificity, and significance issues Public health/regulatory action Practical implications

75 Practical Considerations gent and exposure prevalence information is not generally available Routine real-time 2 nd enzyme assays speeds up investigation and interventions for those situations where it s needed

76 Practical Considerations Multiple case definitions may be needed (e.g. definitions based on 2 enzymes, 1 enzyme, and no PFGE) if the number of cases is small significance specificity of case definition

77 Practical Considerations I you can t use 2 nd enzyme analysis on all recommended pathogens: use for clusters of common subtypes, or when highprevalence exposures are suspected

78 When are 2 nd enzymes likely necessary? Exposure common rare gent common rare ++ + lso recommended High consequence pathogens Multi-state clusters

79 Practical Considerations The 2 enzyme policy is designed to make specific agent data readily available It is NOT a required criteria for epidemiological significance!

80 Practical Considerations Lack of 2 enzyme data is NOT reason to delay a public health action, if a significant association is found with any other case definition

81 Subtype Interpretation subtype mismatch does not automatically exclude association subtype match does not guarantee association The significance of a subtype match is greater if the agent and/or exposure is rare More so than human DN fingerprinting, molecular subtype data cannot be evaluated outside epidemiological context

82 Practical Considerations Obtaining sensitive and specific exposure information is at least as important as agent specificity (Realtime interview, detailed, open-ended questions)

83 Important notes for epidemiology investigators: a Using strict case definitions based on two enzymes is useful for increasing the specificity of hypothesis generation and testing. However, since multiple PFGE strains are on occasion found among epidemiologically-linked cases, investigators should not rule out using both stringent case definitions based on two enzyme results and less stringent case definitions based on single enzyme results. b The significance of clusters is dependent on epidemiological measures. In any particular situation, second enzyme results may or may not ultimately prove to be a useful refinement of the case definition. While it is prudent for laboratories to begin second enzyme testing under these circumstances, this does NOT mean that investigators should wait for the second enzyme results before taking action. c In all cases, investigators should obtain as specific exposure information as possible, which strengthens measures of association between illness and exposure

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