Ananyina 2000.

Study characteristics
Patient sampling	2‐gate case‐control design including people with leptospirosis and people without leptospirosis
Patient characteristics and setting	Sample size: 103 cases, 55 non‐cases (total 158) Clinical presentation: NR for cases; non‐cases had diseases such as acute respiratory infections, hepatitis, lues, Lyme, etc. Risk factors: NR Region: Russia and China, various regions Clinical setting: NR Prevalence of leptospirosis: NA DPO of disease at moment of enrolment: only reported for cases: range 1–5 weeks
Index tests	Index test: PCR Target gene: G and B primers (Gravekamp 1993) Timing of sample collection at # DPO of disease: only reported for cases: range 1–5 weeks 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): acute – see index test; convalescent: NR Single or paired sera (MAT): single and paired Cut‐off value (MAT): NR 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?	No
Did the study avoid inappropriate exclusions?	Unclear
		High	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?	No
Is the execution of the test reported in such detail that reproduction is possible?	No
		Low	High
DOMAIN 3: Reference Standard
Is this the type of test that is likely to correctly classify the target condition?	Yes
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?	No
		Low	High
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