Viromeoffecesfromnewbornnewborn05
Forviralmetagenomicanalysis,90Fecalspecimenswerecollectedfrom45neonateswhowerebornintheAffiliatedHospitalofJiangsuUniversityand45healthychildrenwhowereexaminedatthesamehospitalfromSep.toDec.2018.Eachspecimenwasadded200to500ulDPBS.Andtheneachspecimenwasshakenfor5minutesandrepeateditthreetimes.Thesupernatantswerecollectedaftercentrifugation.Thenthe90supernatantswererandomlypooledinto10specimenpools,eachincluding9specimens.Supernatantswerefilteredthrougha0.45-mmfilter(Millipore)toremoveeukaryotic-andbacterialcell-sizedparticles,and200uLofsupernatantfromeachpoolwasthensubjectedtoamixtureofnucleaseenzymestoreducetheconcentrationoffree(non-viralencapsidated)nucleicacids.RemainingtotalnucleicacidwasthenisolatedusingQIAampMinEluteVirusSpinKitaccordingtomanufacturer'sprotocol.TenlibrarieswerethenconstructedusingNexteraXTDNASamplePreparationKit(Illumina)andsequencedusingtheIlluminasequencingplatformwith250basespairedendswithdualbarcodingforeachpool.