Waggoner 2015.

Study characteristics
Patient sampling	Single‐gate case‐control study with non‐random sampling of participants: up to 55 MAT‐tested samples were selected per month, but if the number exceeded 55, only MAT‐negatives were selected for that month.
Patient characteristics and setting	Sample size: 33 cases, 445 non‐cases (total 478) Clinical presentation: NR Risk factors: NR Region: Brazil, Rio de Janeiro state Clinical setting: NR Prevalence of leptospirosis: NA DPO of disease at moment of enrolment: NR
Index tests	Index test: real‐time PCR (UFI Assay or Lepto‐MD Assay) (TaqMan) Target gene: rrs (Waggoner 2014) Timing of sample collection at # DPO of disease: NR Cut‐off value: 45 Ct Patient material: serum Company/brand: inhouse Frozen samples: yes Note: multiplex real‐time PCR for dengue, plasmodium, and leptospirosis
Target condition and reference standard(s)	Target condition: leptospirosis Case definition: MAT positive Timing of sample collection at # DPO of disease (MAT): NR Single or paired sera (MAT): single Cut‐off value (MAT): ≥ 1:800 Patient material (MAT): serum
Flow and timing	All people were included in the analysis.
Comparative
Notes
Methodological quality
Item	Authors' judgement	Risk of bias	Applicability concerns
DOMAIN 1: Patient Selection
Was a consecutive or random sample of patients enrolled?	No
Was a 2‐gate case‐control design avoided?	Yes
Did the study avoid inappropriate exclusions?	Unclear
		High	Unclear
DOMAIN 2: Index Test Real‐time PCR
Were the index test results interpreted without knowledge of the results of the reference standard?	Unclear
If a threshold was used, was it pre‐specified?	Yes
Does the index test, its conduct and interpretation, match the review question?	No
Is the execution of the test reported in such detail that reproduction is possible?	Yes
		Unclear	High
DOMAIN 3: Reference Standard
Is this the type of test that is likely to correctly classify the target condition?	No
Did the execution of the MAT include paired samples?	Unclear
Were the reference standard results interpreted without knowledge of the results of the index tests?	Yes
Does the case definition match the review question?	Yes
Is the execution of the test reported in such detail that reproduction is possible?	Yes
		High	Low
DOMAIN 4: Flow and Timing
Was there an appropriate interval between index test and reference standard?	Unclear
Did all patients receive a reference standard?	Yes
Did all patients receive the same reference standard?	Yes
Were all patients included in the analysis?	Yes
		Low