Identification of the Pol Gene as a Species-Specific Diagnostic Marker for Qualitative and Quantitative PCR Detection of Tricholoma matsutake

Tricholoma matsutake is a rare, precious, and wild edible fungus that could not be cultivated artificially until now. This situation has given way to the introduction of fake T. matsutake commodities to the mushroom market. Among the methods used to detect food adulteration, amplification of species-specific diagnostic marker is particularly important and accurate. In this study, the Pol gene is reported as a species-specific diagnostic marker to identify three T. matsutake varieties and 10 other types of edible mushrooms through qualitative and quantitative PCR. The PCR results did not reveal variations in the amplified region, and the detection limits of qualitative and quantitative PCR were found to be 8 ng and 32 pg, respectively. Southern blot showed that the Pol gene exists as a single copy in the T. matsutake genome. The method that produced the purest DNA of T. matsutake in this study was also determined, and the high-concentration salt precipitation method was confirmed to be the most suitable among the methods tested. The assay proposed in this work is applicable not only to the detection of raw materials but also to the examination of processed products containing T. matsutake.

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Species-specific identification of ruminant components contaminating industrial crude porcine heparin using real-time fluorescent qualitative and quantitative PCR

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-011-5590-2 ◽

2011 ◽

Vol 402 (4) ◽

pp. 1625-1634 ◽

Cited By ~ 15

Author(s):

Qing Huang ◽

Ting Xu ◽

Gui-Yu Wang ◽

Jun-Fu Huang ◽

Han Xia ◽

...

Keyword(s):

Real Time ◽

Quantitative Pcr ◽

Specific Identification ◽

Qualitative And Quantitative ◽

Qualitative And Quantitative Pcr ◽

Species Specific

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Qualitative and quantitative PCR methods using species-specific primer for detection and identification of wood rot fungi

Journal of Wood Science ◽

10.1007/s10086-008-1011-3 ◽

2009 ◽

Vol 55 (2) ◽

pp. 133-138 ◽

Cited By ~ 15

Author(s):

Sakae Horisawa ◽

Yoh Sakuma ◽

Shuichi Doi

Keyword(s):

Quantitative Pcr ◽

Specific Primer ◽

Qualitative And Quantitative ◽

Detection And Identification ◽

Qualitative And Quantitative Pcr ◽

Wood Rot Fungi ◽

Species Specific

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Simplex and Duplex Polymerase Chain Reaction Analysis of Herculex® RW (59122) Maize Based on One Reference Molecule Including Separated Fragments of 5 Integration Site and Endogenous Gene

Journal of AOAC International ◽

10.1093/jaoac/92.5.1472 ◽

2009 ◽

Vol 92 (5) ◽

pp. 1472-1483 ◽

Cited By ~ 11

Author(s):

Xiang Li ◽

Litao Yang ◽

Jianzhong Zhang ◽

Shu Wang ◽

Kailin Shen ◽

...

Keyword(s):

Quantitative Pcr ◽

Genomic Dna ◽

Raw Materials ◽

Integration Site ◽

Polymerase Chain Reaction Analysis ◽

Detection Methods ◽

Endogenous Gene ◽

Qualitative And Quantitative ◽

Reference Molecule ◽

Qualitative And Quantitative Pcr

Abstract Reference molecules, as positive controls and calibrators, have been recently developed in genetically modified organism analysis as a potential substitute for reference materials derived from plant raw materials. In this study, a novel reference molecule p59122, including the revealed 5 integration sequence of maize Herculex® RW (59122), was constructed that was suitable for simplex and duplex event-specific qualitative and quantitative PCR detections. The LOD values were 10 copies both for simplex and duplex qualitative PCR when p59122 was used as the calibrator. These values were comparable to those of using genomic DNA samples with 0.01 and 0.05, approximately 5 and 25 hyploid genomic DNA copies, respectively. The absolute LOD and LOQ values were confirmed to be as low as 10 and 25 copies of p59122 DNA both in simplex and duplex quantitative systems. Furthermore, ideal quantification data with low bias, SD and RSD values were obtained from the practical samples analyses in simplex and duplex real-time PCR systems using the reference molecule p59122 as a calibrator. All these results suggested that the developed reference molecule p59122 and the qualitative and quantitative PCR detection methods are suitable for identification and quantification of GM maize 59122 and its derived products.

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