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Collaborative Ring Trial of the Papaya Endogenous Reference Gene and Its Polymerase Chain Reaction Assays for Genetically Modified Organism Analysis

文献类型： 外文期刊

作者： Wei, Jiaojun 1 ; Li, Feiwu 2 ; Guo, Jinchao 1 ; Li, Xiang 3 ; Xu, Junfeng 4 ; Wu, Gang 5 ; Zhang, Dabing 1 ; Yang, Litao 1 ;

作者机构： 1.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, SJTU Bor Luh Food Safety Ctr, Natl Ctr Mol Characterizat Genetically Modified O, Shanghai 200240, Peoples R China

2.Jilin Acad Agr Sci, Agrobiotechnol Res Inst, Changchun 130033, Peoples R China

3.Shanghai Entry Exit Inspect & Quarantine Bur, GMO Detect Lab, Shanghai 200135, Peoples R China

4.Zhejiang Acad Agr Sci, Inst Agr Qual & Stand Agroprod, Hangzhou 310021, Zhejiang, Peoples R China

5.Chinese Acad Agr Sci, Oil Crops Res Inst, Minist Agr, Key Lab Oil Crop Biol, Wuha

关键词： collaborative ring trial;Carica papaya;Chymopapain;endogenous reference gene;qualitative and quantitative PCR

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.279； 五年影响因子：5.269 ）

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收录情况： SCI

摘要： The papaya (Carka papaya L.) Chymopapain (CHY) gene has been reported as a suitable endogenous reference gene for genetically modified (GM) papaya detection in previous studies. Herein, we further validated the use of the CHY gene and its qualitative and quantitative polymerase chain reaction (PCR)' assays through an interlaboratory collaborative ring trial. A total of 12 laboratories working on detection of genetically modified organisms participated in the ring trial and returned test results. Statistical analysis of the returned results confirmed the species specificity, low heterogeneity, and single-copy number of the CHY gene among different papaya varieties. The limit of detection of the CHY qualitative PCR assay was 0.1%, while the limit of quantification of the quantitative PCR assay was ～25 copies of haploid papaya genome with acceptable PCR efficiency and linearity. The differences between the tested and true values of papaya content in 10 blind samples ranged from 0.84 to 6.58%. These results indicated that the CHY gene was suitable as an endogenous reference gene for the identification and quantification of GM papaya.

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