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Duplex real-time PCR for sexing Schistosoma japonicum cercariae based on W chromosome-specific genes and its applications.
- Source :
-
PLoS neglected tropical diseases [PLoS Negl Trop Dis] 2020 Aug 21; Vol. 14 (8), pp. e0008609. Date of Electronic Publication: 2020 Aug 21 (Print Publication: 2020). - Publication Year :
- 2020
-
Abstract
- As a unique feature among otherwise hermaphroditic trematodes, Schistosoma species are gonochoric parasites whose sex is genetically determined (ZZ for males and ZW for females). However, schistosome larvae are morphologically identical, and sex can only be discriminated by molecular methods. Here, we integrated published Schistosoma. japonicum transcriptome and genome data to identify W chromosome-specific genes as sex biomarkers. Three W chromosome-specific genes of S. japonicum were identified as sex biomarkers from a panel of 12 genes expressed only in females. An efficient duplex real-time PCR (qPCR) method for sexing cercariae was developed which could identify the sex of cercariae within 2 h without DNA extraction. Moreover, this method can be used to identify not only single-sex but also mixed-sex schistosome-infected snails. We observed a nearly equal proportion of single-male, single-female, and mixed-sex schistosome infections in artificially infected snails. Sex-known schistosome-infected snail models can be efficiently constructed with the aid of duplex qPCR. A field study revealed that single-sex schistosome infections were predominant among naturally infected snails. Finally, a schistosomiasis mouse model based on sex-known cercariae infection was shown to be more reliable than a model based on sex-unknown cercariae infection. The developed duplex qPCR method for sexing S. japonicum cercariae can be widely used for schistosomiasis modeling, genetic experiments, and field-based molecular epidemiological studies.<br />Competing Interests: The authors have declared that no competing interests exist.
- Subjects :
- Animals
Biomarkers
China epidemiology
DNA isolation & purification
Disease Models, Animal
Female
Gene Expression
Male
Mice
Mice, Inbred BALB C
Schistosomiasis epidemiology
Snails parasitology
Cercaria genetics
Chromosomes
Real-Time Polymerase Chain Reaction methods
Schistosoma japonicum genetics
Schistosomiasis parasitology
Subjects
Details
- Language :
- English
- ISSN :
- 1935-2735
- Volume :
- 14
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- PLoS neglected tropical diseases
- Publication Type :
- Academic Journal
- Accession number :
- 32822351
- Full Text :
- https://doi.org/10.1371/journal.pntd.0008609