Objectives:Cronobacter sakazakii,formerly Enterobacter sakazakii,is an emerging ubiquitous and opportunistic foodborne pathogen with a high mortality rate.It has been implicated in cases of meningitis,septicaemia,and ...Objectives:Cronobacter sakazakii,formerly Enterobacter sakazakii,is an emerging ubiquitous and opportunistic foodborne pathogen with a high mortality rate.It has been implicated in cases of meningitis,septicaemia,and necrotizing enterocolitis among infants worldwide in association with powdered infant formula(PIF).This study was an insilico designed peptide base kit framework,using immunoinformatic techniques for quick detection of C.sakazakii in PIF.Materials and Methods:In the present study,a peptide-based kit was designed with a bioinformatic technique to rapidly identify C.sakazakii in PIF using fhE,secY,and bcsC,which are genes responsible for its bioflm formation,as target genes.The antigenicity,membrane topology,and the presence of signal peptides of the target genes were analysed using VaxiJen,DeepTMHMM,and SignalP servers.To provide stability and fexibility to the multiple-epitope construct,the linear B cells and helper T cells(IL-4(interleukin 4)and IL-10(interleukin 10)inducing epitopes)were linked with a GSGSG linker followed by the addition of protein disulphide bonds.To ascertain specifcity,the multi-epitope construct was molecularly docked against genes from sources other than PIF,like alfalfa,and the environment,with PIF being the highest:–328.48.Finally,the codons were modifed using the pET28a(+)vector,and the resultant multi-epitope construct was successfully cloned in silico.Results:The fnal construct had a length of 486 bp,an instability index of 23.26,a theoretical pI of 9.34,a molecular weight of 16.5 kDa,and a Z-score of–3.41.Conclusions:The multi-epitope peptide construct could be a conceptual framework for creating a C.sakazakii peptide-based detection kit,which has the potential to provide fast and effcient detection.However,there is a need for additional validation through the in vitro and in vivo techniques.展开更多
文摘Objectives:Cronobacter sakazakii,formerly Enterobacter sakazakii,is an emerging ubiquitous and opportunistic foodborne pathogen with a high mortality rate.It has been implicated in cases of meningitis,septicaemia,and necrotizing enterocolitis among infants worldwide in association with powdered infant formula(PIF).This study was an insilico designed peptide base kit framework,using immunoinformatic techniques for quick detection of C.sakazakii in PIF.Materials and Methods:In the present study,a peptide-based kit was designed with a bioinformatic technique to rapidly identify C.sakazakii in PIF using fhE,secY,and bcsC,which are genes responsible for its bioflm formation,as target genes.The antigenicity,membrane topology,and the presence of signal peptides of the target genes were analysed using VaxiJen,DeepTMHMM,and SignalP servers.To provide stability and fexibility to the multiple-epitope construct,the linear B cells and helper T cells(IL-4(interleukin 4)and IL-10(interleukin 10)inducing epitopes)were linked with a GSGSG linker followed by the addition of protein disulphide bonds.To ascertain specifcity,the multi-epitope construct was molecularly docked against genes from sources other than PIF,like alfalfa,and the environment,with PIF being the highest:–328.48.Finally,the codons were modifed using the pET28a(+)vector,and the resultant multi-epitope construct was successfully cloned in silico.Results:The fnal construct had a length of 486 bp,an instability index of 23.26,a theoretical pI of 9.34,a molecular weight of 16.5 kDa,and a Z-score of–3.41.Conclusions:The multi-epitope peptide construct could be a conceptual framework for creating a C.sakazakii peptide-based detection kit,which has the potential to provide fast and effcient detection.However,there is a need for additional validation through the in vitro and in vivo techniques.
