Biscornet 2017.

Study characteristics
Patient sampling	Cross‐sectional study with consecutive sampling method.
Patient characteristics and setting	Sample size: 31 cases, 192 non‐cases (total 223) Clinical presentation: acute fever of unknown origin > 3 days, with or without any of the following: headaches, myalgia, haemorrhagic manifestations in the absence of any definite diagnosis. Risk factors: NR Region: Seychelles, nationwide Clinical setting: population‐based national survey Prevalence of leptospirosis: 13.9% DPO of disease at moment of enrolment: NR
Index tests	Index test: real‐time PCR (TaqMan) Target gene: rrs (Smythe 2002) Timing of sample collection at # DPO of disease: NR Cut‐off value: 35 Ct Patient material: serum Company/brand: inhouse Frozen samples: yes
Target condition and reference standard(s)	Target condition: leptospirosis Case definition: MAT or IgM ELISA positive, or both Timing of sample collection at # DPO of disease (MAT): acute: NR; convalescent: ≥ 4 weeks DPO Single or paired sera (MAT): single and paired Cut‐off value (MAT): 1:400 or 4‐fold titre rise Patient material (MAT): serum Timing of sample collection at # DPO of disease (IgM ELISA): acute: NR; convalescent: ≥ 4 weeks DPO Single or paired sera (IgM ELISA): NR Cut‐off value (IgM ELISA): NR Patient material (IgM ELISA): 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?	Yes
Was a 2‐gate case‐control design avoided?	Yes
Did the study avoid inappropriate exclusions?	Yes
		Low	Low
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?	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?	Unclear
Does the case definition match the review question?	Yes
Is the execution of the test reported in such detail that reproduction is possible?	Yes
		Unclear	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