Domain	Participant selection	Index test	Reference standard	Flow and timing
Description	Describe methods of participant selection: describe included participant's (prior testing, presentation, intended use of index test and setting)	Describe the index test and how it was conducted and interpreted (where the laboratory test was applied, by whom, who interpreted the results of the test, what was considered a positive result)	Describe the reference standard and how it was conducted and interpreted (what is the reference standard used, definition of positive result, who interpreted the results of the reference standard, etc)	Describe any participants who did not receive the index test(s) or reference standard, or both, or who were excluded from the 2 × 2 table (refer to flow diagram): describe the time interval and any interventions between index test(s) and reference standard
Signalling questions (yes/no/unclear)	Was a consecutive or random sample of neonates enrolled? Yes = a consecutive or random sample of neonates was enrolled Unclear = insufficient information provided No = selected sample of neonates were analysed	Were the congenital Zika virus test results interpreted without knowledge of the results of the reference standard? Yes = the index test was interpreted without knowledge of the reference standard findings Unclear = insufficient information provided No = the index test was interpreted with knowledge of the reference standard findings, or the index test was part of the reference standard	Is the reference standard likely to correctly classify the target condition? Yes = a reference standard from those listed in Methods was applied Unclear = insufficient information provided No = other criteria, even isolated findings, were applied to diagnose CZV	Was there an appropriate interval between index test(s) and reference standard? Yes = the index test and the reference standard are administered within 2 weeks after birth, or at the same time Unclear = insufficient information provided No = the index test and the reference standard were administered in different times after birth (> 2 weeks)
	Did the study avoid inappropriate exclusions? Yes = all neonates who were candidates to be tested were included Unclear = insufficient information provided No = the study excluded neonates who were candidates for testing	If a threshold was used, was it prespecified? Yes = the positivity threshold was established before the administration of the index test Unclear = insufficient information provided No = the positivity threshold was not established after the administration of the index test results (i.e. optimal cut‐off)	Were the reference standard results interpreted without knowledge of the results of the index test? Yes = the reference standard was interpreted without knowledge of the index test results Unclear = insufficient information provided No = the reference standard was interpreted with knowledge of the index test results, or the index test was part of the reference standard	Did all participants receive a reference standard? Yes = a reference standard was applied regardless of the index test results Unclear = insufficient information provided No = only a sample of neonates received the reference standard (for example, those with positive index test results)
				Did all participants receive the same reference standard? Yes = a unique reference standard was applied, regardless of the index test results Unclear = insufficient information provided No = more than one reference standard was applied, mostly depending on index test results
				Were all participants included in the analysis? Yes= All neonates who were enrolled were analysed Unclear = insufficient information provided No = the study excluded participants from final analysis (i.e. indeterminate results)
Risk of bias (high/low/unclear)	Could the selection of participants have introduced bias?	Could the conduct or interpretation of the index test have introduced bias?	Could the reference standard, its conduct, or its interpretation have introduced bias?	Could the participant flow have introduced bias?
Concerns regarding applicability (high/low/unclear)	Are there concerns that the included participants did not match the review question?	Are there concerns that the index test, its conduct, or interpretation differed from the review question?	Are there concerns that the target condition, as defined by the reference standard, did not match the review question?