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Analysis on Gene Expression Profile in Oncospheres and Early Stage Metacestodes from Echinococcus multilocularis.

Huang F, Dang Z, Suzuki Y, Horiuchi T, Yagi K, Kouguchi H, Irie T, Kim K, Oku Y - PLoS Negl Trop Dis (2016)

Bottom Line: However, little gene expression data is available for antigens of the egg and early larval stages.Furthermore, heat shock proteins and antigen II/3 are constantly expressed in the three stages.The expression pattern of various known antigens in E. multilocularis may give fundamental information for choosing candidate genes used in diagnosis and vaccine development.

View Article: PubMed Central - PubMed

Affiliation: Parasitology Laboratory, School of Veterinary Medicine, Faculty of Agriculture, Tottori University, Tottori, Japan.

ABSTRACT
Alveolar echinococcosis is a worldwide zoonosis of great public health concern. Analysis of genome data for Echinococcus multilocularis has identified antigen families that can be used in diagnostic assays and vaccine development. However, little gene expression data is available for antigens of the egg and early larval stages. To address this information gap, we used a Next-Generation Sequencing approach to investigate three different stages (non-activated and activated oncospheres, and early stage metacestodes) of E. multilocularis (Nemuro strain). Transcriptome data analysis revealed that some diagnostic antigen gp50 isoforms and the antigen Eg95 family dominated in activated oncospheres, and the antigen B family dominated in early stage metacestodes. Furthermore, heat shock proteins and antigen II/3 are constantly expressed in the three stages. The expression pattern of various known antigens in E. multilocularis may give fundamental information for choosing candidate genes used in diagnosis and vaccine development.

No MeSH data available.


Related in: MedlinePlus

Real-time PCR validation of genes at kinds of expression level.The y-axis indicates the value of relative expression level (2-ΔΔCt) by real-time PCR and log2Ratio of Nonc1/Cmet by Next-generation sequencing. GAPDH as the internal control. Error Bar: Standard Deviation of the mean expression values of Real-time PCR.
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pntd.0004634.g002: Real-time PCR validation of genes at kinds of expression level.The y-axis indicates the value of relative expression level (2-ΔΔCt) by real-time PCR and log2Ratio of Nonc1/Cmet by Next-generation sequencing. GAPDH as the internal control. Error Bar: Standard Deviation of the mean expression values of Real-time PCR.

Mentions: We constructed five cDNA libraries from Nonc1, Nonc2, Aonc, 4Wmet and Cmet of E. multilocularis (Fig 1). More than 493 million clean reads were generated by Illumina paired-end sequencing, and 9,852 coding sequences of the genome were mapped with RPKM bigger than zero in at least one of the sequenced samples (S2 Table). The quality of obtained reads was excellent with more than 90% of reads having a quality score at Q30 (error probability of 0.001) or higher (Table 2). The results of real-time PCR analysis confirmed the NGS analysis data and show similar trends in fold change (Fig 2).


Analysis on Gene Expression Profile in Oncospheres and Early Stage Metacestodes from Echinococcus multilocularis.

Huang F, Dang Z, Suzuki Y, Horiuchi T, Yagi K, Kouguchi H, Irie T, Kim K, Oku Y - PLoS Negl Trop Dis (2016)

Real-time PCR validation of genes at kinds of expression level.The y-axis indicates the value of relative expression level (2-ΔΔCt) by real-time PCR and log2Ratio of Nonc1/Cmet by Next-generation sequencing. GAPDH as the internal control. Error Bar: Standard Deviation of the mean expression values of Real-time PCR.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836691&req=5

pntd.0004634.g002: Real-time PCR validation of genes at kinds of expression level.The y-axis indicates the value of relative expression level (2-ΔΔCt) by real-time PCR and log2Ratio of Nonc1/Cmet by Next-generation sequencing. GAPDH as the internal control. Error Bar: Standard Deviation of the mean expression values of Real-time PCR.
Mentions: We constructed five cDNA libraries from Nonc1, Nonc2, Aonc, 4Wmet and Cmet of E. multilocularis (Fig 1). More than 493 million clean reads were generated by Illumina paired-end sequencing, and 9,852 coding sequences of the genome were mapped with RPKM bigger than zero in at least one of the sequenced samples (S2 Table). The quality of obtained reads was excellent with more than 90% of reads having a quality score at Q30 (error probability of 0.001) or higher (Table 2). The results of real-time PCR analysis confirmed the NGS analysis data and show similar trends in fold change (Fig 2).

Bottom Line: However, little gene expression data is available for antigens of the egg and early larval stages.Furthermore, heat shock proteins and antigen II/3 are constantly expressed in the three stages.The expression pattern of various known antigens in E. multilocularis may give fundamental information for choosing candidate genes used in diagnosis and vaccine development.

View Article: PubMed Central - PubMed

Affiliation: Parasitology Laboratory, School of Veterinary Medicine, Faculty of Agriculture, Tottori University, Tottori, Japan.

ABSTRACT
Alveolar echinococcosis is a worldwide zoonosis of great public health concern. Analysis of genome data for Echinococcus multilocularis has identified antigen families that can be used in diagnostic assays and vaccine development. However, little gene expression data is available for antigens of the egg and early larval stages. To address this information gap, we used a Next-Generation Sequencing approach to investigate three different stages (non-activated and activated oncospheres, and early stage metacestodes) of E. multilocularis (Nemuro strain). Transcriptome data analysis revealed that some diagnostic antigen gp50 isoforms and the antigen Eg95 family dominated in activated oncospheres, and the antigen B family dominated in early stage metacestodes. Furthermore, heat shock proteins and antigen II/3 are constantly expressed in the three stages. The expression pattern of various known antigens in E. multilocularis may give fundamental information for choosing candidate genes used in diagnosis and vaccine development.

No MeSH data available.


Related in: MedlinePlus