Interference of Real-Time PCR Quantification of<i>Vibrio vulnificus</i>by a Novel DNase from the Eastern Oyster (<i>Crassostrea virginica</i>)

Wang, Shishan; Levin, Robert E.

doi:10.1080/08905431003784721

Cited by 3 publications

(1 citation statement)

References 26 publications

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“…Another major problem with detecting pathogenic bacteria in shellfish tissue without enrichment is the high level of shellfish DNase present in the tissue which must be removed before cell lysis. These problems have been previously addressed [39,40].…”

Section: Resultsmentioning

confidence: 99%

http://www.omicsonline.org/open-access/overcoming-pseudomonas-aeruginosa-resistance-caused-by-glycocalyx-with-tobracef-1948-5948.1000145.php?aid=25925

Dai¹

2014

J Microb Biochem Technol

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Oysters are filter feeders that bioaccumulate bacteria in water while feeding. To evaluate the bacterial genomic DNA extracted from retail oyster tissues, including the gills and digestive glands, four isolation methods were used. Genomic DNA extraction was performed using the Allmag™ Blood Genomic DNA (Allrun, Shanghai, China), the MiniBEST Bacterial Genomic DNA Extraction kits (Takara, Dalian, China), and the phenol-chloroform and boiling lysis methods. The concentration of the genomic DNA was measured using a spectrophotometer. The purity of the genomic DNA was evaluated by PCR amplification of 16S rDNA followed by determining the cloning efficiency of the amplicon into the pMD19-T vector. Furthermore, the bacterial DNA quality was also evaluated by PCR assays using a pair of species-specific primers for Vibrio parahaemolyticus. Our results showed that the two commercial kits produced the highest purity of DNA, but with the lowest yields. The phenol-chloroform method produced the highest yield although it was time-consuming. The boiling lysis method was simple and cost effective; however, it was only suitable to isolate genomic DNA from bacteria present in retail samples following an enrichment step. The two commercial kits were good candidates for genomic DNA extraction from retail oyster tissues without enrichment.

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Section: Resultsmentioning

confidence: 99%

http://www.omicsonline.org/open-access/overcoming-pseudomonas-aeruginosa-resistance-caused-by-glycocalyx-with-tobracef-1948-5948.1000145.php?aid=25925

Dai¹

2014

J Microb Biochem Technol

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Characterization of a Recently Purified Thermophilic DNase from a Novel Thermophilic Fungus

Landry

Levin

2014

Appl Biochem Biotechnol

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A newly isolated thermophilic fungus was found to produce a partially inducible extracellular DNase. This manuscript focuses on the characterization of this novel thermophilic DNase in terms of optimal enzyme conditions, molecular weight, and certain kinetic properties. The DNase was found to be inactivated by the presence of EDTA demonstrating its dependence on metal cofactors for activity. Maximum activity occurred at pH 6.0 with no activity at pH 2.0 or 10.0. The optimal temperature for the purified DNase was 65 °C. The thermophilic DNase was found to be an exonuclease with an estimated molecular weight of 56 kDa.

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Recent Developments in Molecular Detection of Food-Borne Disease Bacteria and GMOs

Levin

2015

Food Biotechnology

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Interference of Real-Time PCR Quantification ofVibrio vulnificusby a Novel DNase from the Eastern Oyster (Crassostrea virginica)

Cited by 3 publications

References 26 publications

http://www.omicsonline.org/open-access/overcoming-pseudomonas-aeruginosa-resistance-caused-by-glycocalyx-with-tobracef-1948-5948.1000145.php?aid=25925

http://www.omicsonline.org/open-access/overcoming-pseudomonas-aeruginosa-resistance-caused-by-glycocalyx-with-tobracef-1948-5948.1000145.php?aid=25925

Characterization of a Recently Purified Thermophilic DNase from a Novel Thermophilic Fungus

Recent Developments in Molecular Detection of Food-Borne Disease Bacteria and GMOs

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