Triplex real-time PCR–an improved method to detect a wide spectrum of mitochondrial DNA deletions in single cells

A sensitive and rapid method for the quantitative detection of bovine-, ovine-, swine-, and chicken-specific mitochondrial DNA sequences based on real-time PCR has been developed. The specificity of the primers and probes for real-time PCR has been tested using DNA samples of other vertebrate species that may also be present in rendered products. The quantitative detection was performed with dual-labeled probes (TaqMan) using absolute quantification with external standards of single species meat-and-bone meals. This method facilitates the detection of 0.01% of the target species–derived material in concentrate feed mixtures and fish meals.

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Real-Time PCR Detection of Ruminant DNA

Journal of Food Protection ◽

10.4315/0362-028x-67.3.550 ◽

2004 ◽

Vol 67 (3) ◽

pp. 550-554 ◽

Cited By ~ 36

Author(s):

LUIS MENDOZA-ROMERO ◽

EDWARD L. C. VERKAAR ◽

PAUL H. SAVELKOUL ◽

ARNOLD CATSBURG ◽

HENK J. M. AARTS ◽

...

Keyword(s):

Mitochondrial Dna ◽

Bovine Spongiform Encephalopathy ◽

Real Time ◽

Real Time Pcr ◽

Spongiform Encephalopathy ◽

Meat And Bone Meal ◽

Bone Meal ◽

Dna Targeting ◽

Species Origin ◽

Semiquantitative Method

To control the spread of bovine spongiform encephalopathy, several DNA methods have been described for the detection of the species origin of meat and bone meal. Most of these methods are based on the amplification of a mitochondrial DNA segment. We have developed a semiquantitative method based on real-time PCR for detection of ruminant DNA, targeting an 88-bp segment of the ruminant short interspersed nuclear element Bov-A2. This method is specific for ruminants and is able to detect as little as 10 fg of bovine DNA. Autoclaving decreased the amount of detectable DNA, but positive signals were observed in feeding stuff containing 10% bovine material if this had not been rendered in accordance with the regulations, i.e., heated at 134°C for 3 instead of 20 min.

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Interlaboratory validation of an improved method for detection of Cyclospora cayetanensis in produce using a real-time PCR assay

Food Microbiology ◽

10.1016/j.fm.2017.08.008 ◽

2018 ◽

Vol 69 ◽

pp. 170-178 ◽

Cited By ~ 14

Author(s):

Helen R. Murphy ◽

Hediye Nese Cinar ◽

Gopal Gopinath ◽

Kathy E. Noe ◽

Lacresha D. Chatman ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Improved Method ◽

Pcr Assay ◽

Cyclospora Cayetanensis ◽

Interlaboratory Validation

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Common Deletion (CD) in mitochondrial DNA of irradiated rat heart

Anais da Academia Brasileira de Ciências ◽

10.1590/0001-37652014110912 ◽

2014 ◽

Vol 86 (2) ◽

pp. 685-694 ◽

Cited By ~ 1

Author(s):

RAQUEL G. SIQUEIRA ◽

DAYSE A. DA SILVA ◽

LUIZ D.B. DE MELO ◽

ELIZEU F. DE CARVALHO ◽

SAMARA C. FERREIRA-MACHADO ◽

...

Keyword(s):

Mitochondrial Dna ◽

Ionizing Radiation ◽

Real Time ◽

Real Time Pcr ◽

Rat Heart ◽

Wistar Rats ◽

Post Irradiation ◽

Male Wistar Rats ◽

The Common ◽

Common Deletion

The purpose of this study was to map the common deletion (CD) area in mtDNA and investigate the levels of this deletion in irradiated heart. The assays were developed in male Wistar rats that were irradiated with three different single doses (5, 10 or 15 Gy) delivered directly to the heart and the analyses were performed at various times post-irradiation (3, 15 or 120 days). The CDs area were sequenced and the CD quantified by real-time PCR. Our study demonstrated that the CD levels progressively decreased from the 3rduntil the 15th day after irradiation, and then increased thereafter. Additionally, it was observed that the levels of CD are modulated differently according to the different categories of doses (moderate and high). This study demonstrated an immediate response to ionizing radiation, measured by the presence of mutations in the CD area and a decrease in the CD levels.

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