Mitochondrial genome sequence variation as a useful marker for assessing genetic heterogeneity among Cyclospora cayetanensis isolates and source-tracking.
Cyclospora cayetanensis
Genotyping
Mitochondrion
Source-tracking
qPCR
Journal
Parasites & vectors
ISSN: 1756-3305
Titre abrégé: Parasit Vectors
Pays: England
ID NLM: 101462774
Informations de publication
Date de publication:
21 Jan 2019
21 Jan 2019
Historique:
received:
30
10
2018
accepted:
07
01
2019
entrez:
23
1
2019
pubmed:
23
1
2019
medline:
29
1
2019
Statut:
epublish
Résumé
Cyclospora cayetanensis is an important enteric pathogen, causing diarrhea and food-borne cyclosporiasis outbreaks. For effective outbreak identification and investigation, it is essential to rapidly assess the genetic heterogeneity of C. cayetanensis specimens from cluster cases and identify the likely occurrence of outbreaks. In this study, we developed a quantitative PCR (qPCR) targeting the polymorphic link region between copies of the mitochondrial genome of C. cayetanensis, and evaluated the genetic heterogeneity among 36 specimens from six countries using melt curve, gel electrophoresis, and sequence analyses of the qPCR products. All specimens were amplified successfully in the qPCR and produced melt peaks with different Tm values in the melt curve analysis. In gel electrophoresis of the qPCR products, the specimens yielded bands of variable sizes. Nine genotypes were identified by DNA sequencing of the qPCR products. Geographical segregation of genotypes was observed among specimens analyzed, which could be useful in geographical source-tracking. The length and nucleotide sequence variations in the mitochondrial genome marker allow rapid assessment of the genetic heterogeneity among C. cayetanensis specimens by melt curve, gel electrophoresis, or DNA sequence analysis of qPCR products. The sequence data generated could be helpful in the initial source-tracking of the pathogen.
Sections du résumé
BACKGROUND
BACKGROUND
Cyclospora cayetanensis is an important enteric pathogen, causing diarrhea and food-borne cyclosporiasis outbreaks. For effective outbreak identification and investigation, it is essential to rapidly assess the genetic heterogeneity of C. cayetanensis specimens from cluster cases and identify the likely occurrence of outbreaks.
METHODS
METHODS
In this study, we developed a quantitative PCR (qPCR) targeting the polymorphic link region between copies of the mitochondrial genome of C. cayetanensis, and evaluated the genetic heterogeneity among 36 specimens from six countries using melt curve, gel electrophoresis, and sequence analyses of the qPCR products.
RESULTS
RESULTS
All specimens were amplified successfully in the qPCR and produced melt peaks with different Tm values in the melt curve analysis. In gel electrophoresis of the qPCR products, the specimens yielded bands of variable sizes. Nine genotypes were identified by DNA sequencing of the qPCR products. Geographical segregation of genotypes was observed among specimens analyzed, which could be useful in geographical source-tracking.
CONCLUSIONS
CONCLUSIONS
The length and nucleotide sequence variations in the mitochondrial genome marker allow rapid assessment of the genetic heterogeneity among C. cayetanensis specimens by melt curve, gel electrophoresis, or DNA sequence analysis of qPCR products. The sequence data generated could be helpful in the initial source-tracking of the pathogen.
Identifiants
pubmed: 30665345
doi: 10.1186/s13071-019-3294-1
pii: 10.1186/s13071-019-3294-1
pmc: PMC6341762
doi:
Substances chimiques
Biomarkers
0
DNA, Protozoan
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
47Subventions
Organisme : National Key Research and Development Program of China
ID : 2017YFD0500400
Organisme : National Natural Science Foundation of China
ID : 31630078
Organisme : National Natural Science Foundation of China
ID : 31425025
Références
Int J Parasitol. 2000 Apr 10;30(4):371-90
pubmed: 10731561
Emerg Infect Dis. 2008 Oct;14(10):1567-74
pubmed: 18826821
Clin Microbiol Rev. 2010 Jan;23(1):218-34
pubmed: 20065331
Acta Trop. 2010 Sep;115(3):181-93
pubmed: 20382099
Emerg Infect Dis. 2011 Oct;17(10):1887-90
pubmed: 22000362
Korean J Parasitol. 2012 Mar;50(1):57-62
pubmed: 22451735
Curr Opin Infect Dis. 2013 Oct;26(5):479-83
pubmed: 23982239
Am J Trop Med Hyg. 2014 Jun;90(6):1102-8
pubmed: 24639297
Nat Commun. 2014 Jun 13;5:4052
pubmed: 24923250
Int J Parasitol. 2015 May;45(6):361-5
pubmed: 25812835
Epidemiol Infect. 2015 Dec;143(16):3451-8
pubmed: 25865140
Parasit Vectors. 2015 Jul 08;8:358
pubmed: 26152563
Epidemiol Infect. 2016 Jul;144(9):1803-6
pubmed: 26489478
Genome Announc. 2015 Nov 19;3(6):null
pubmed: 26586880
BMC Genomics. 2016 Apr 30;17:316
pubmed: 27129308
Emerg Infect Dis. 2016 Aug;22(8):1464-7
pubmed: 27433881
J Food Prot. 2017 Oct;80(10):1666-1669
pubmed: 28876133
Exp Parasitol. 2018 Jan;184:121-127
pubmed: 29274312
Gut Pathog. 2018 Apr 10;10:15
pubmed: 29643938