Blanco 2014.

Study characteristics
Patient sampling	Cross‐sectional design with unclear sampling method.
Patient characteristics and setting	Sample size: 28 cases, 493 non‐cases (total 521) Clinical presentation: NR ("suspected leptospirosis") Risk factors: NR Region: Brazil Clinical setting: NR Prevalence of leptospirosis: 5.4% DPO of disease at moment of enrolment: NR
Index tests	Index test 1: PCR Target gene: rrs (Merien 1992) Timing of sample collection at # DPO of disease: only reported for cases: 1–10 DPO Cut‐off value: not applicable Patient material: serum Company/brand: inhouse Frozen samples: unclear Index test 2: nested PCR Target gene: rrs (Merien 1992) Timing of sample collection at # DPO of disease: only reported for cases: 1–10 DPO Cut‐off value: not applicable Patient material: serum Company/brand: inhouse Frozen samples: unclear
Target condition and reference standard(s)	Target condition: leptospirosis Case definition: MAT positive Timing of sample collection at # DPO of disease (MAT): only reported for cases – acute: see index test; convalescent: 11–17 DPO. Single or paired sera (MAT): single and paired Cut‐off value (MAT): ≥ 1:800, seroconversion, or a 4‐fold titre rise 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?	Unclear
Was a 2‐gate case‐control design avoided?	Yes
Did the study avoid inappropriate exclusions?	Unclear
		Unclear	Unclear
DOMAIN 2: Index Test PCR
Were the index test results interpreted without knowledge of the results of the reference standard?	Yes
If a threshold was used, was it pre‐specified?	Yes
Does the index test, its conduct and interpretation, match the review question?	Yes
Is the execution of the test reported in such detail that reproduction is possible?	Yes
		Low	Low
DOMAIN 2: Index Test Nested PCR
Were the index test results interpreted without knowledge of the results of the reference standard?	Yes
If a threshold was used, was it pre‐specified?	Yes
Does the index test, its conduct and interpretation, match the review question?	Yes
Is the execution of the test reported in such detail that reproduction is possible?	Yes
		Low	Low
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?	Yes
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