Pakoa 2018.

Study characteristics
Patient sampling	Cross‐sectional study with unclear sampling method.
Patient characteristics and setting	Sample size: 15 cases, 115 non‐cases (total 130) Clinical presentation: 'Feeling unwell', headache, fever or chills, myalgia or arthralgia, prostration, cough, jaundice, and oliguria or anuria Risk factors: NR Region: Port Vila, Vanuatu (island of Efate) Clinical setting: outpatient clinic at a hospital Prevalence of leptospirosis: 11.5% DPO of disease at moment of enrolment: NR
Index tests	Index test: real‐time PCR (TaqMan) Target gene: lipL32 (Stoddard 2009) Timing of sample collection at # DPO of disease: NR Cut‐off value: NR 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): NR Single or paired sera (MAT): single Cut‐off value (MAT): ≥ 1:800 Patient material (MAT): serum
Flow and timing	31 people not included because the amount of serum was not enough for MAT.
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	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?	No
Does the index test, its conduct and interpretation, match the review question?	Unclear
Is the execution of the test reported in such detail that reproduction is possible?	No
		High	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?	No
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
		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?	No
Did all patients receive the same reference standard?	Yes
Were all patients included in the analysis?	No
		High