[Screening and Applications of qPCR Primers for apomucin Gene of Echinococcus multilocularis].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
; 34(5): 394-8, 2016 Oct.
Article
em Zh
| MEDLINE
| ID: mdl-30130005
ABSTRACT
Objective:
To screen for the optimal qPCR primers for Echinococcus multilocularis apomucin gene (Em-apo) and analyze Em-apo expression.Methods:
Primers were designed based on 4 Em-apo sequences from GeneDB. Primer specificity and PCR efficiency were determined, based on which the optimal primer pairs were selected. Alterations of Em-apo expression in 1 000 E. multilocularis protoscoleces treated with albendazoleï¼5 µg/mlï¼ and insulinï¼100 ng/mlï¼ were separately assessed using the selected primers. DMSO used in albendazole dilution and in PBS insulin dilution were used as the control.Results:
Specific primers for Em-apo-1, Em-apo-2/3, Em-apo-4 and actin were selected. qPCR melting curves revealed a single peak for each primer pair and an amplification efficiency from 95% to 101%. The qPCR showed increased expression of Em-apo-1ï¼1.51±0.27ï¼, Em-apo-2/3 ï¼1.39±0.30ï¼ and Em-apo-4ï¼1.14±0.18ï¼ after albendazole treatment in comparison to the DMSO controlï¼1.00ï¼ï¼P>0.05 among the three genesï¼; and an unaltered Em-apo-1 expression, slightly decreased Em-apo-4 expression, and significantly decreased Em-apo-2/3 expressionï¼0.73±0.09ï¼ after insulin treatment in comparison to the PBS control (P>0.05 among the three genesï¼.Conclusion:
The selected specific primers for Em-apo genes can be used to analyze the gene expression by qPCR. Treatment with albendazole and insulin show certain effects on the expression of Em-apo genes in E. multilocularis protoscoleces.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Echinococcus multilocularis
Tipo de estudo:
Diagnostic_studies
/
Screening_studies
Limite:
Animals
Idioma:
Zh
Revista:
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
Assunto da revista:
PARASITOLOGIA
Ano de publicação:
2016
Tipo de documento:
Article