A new quantitative PCR assay for the detection of hepatotoxigenic cyanobacteria

Al-Tebrineh, Jamal; Gehringer, Michelle M.; Akçaalan, Reyhan; Neilan, Brett A.

doi:10.1016/j.toxicon.2010.12.018

Cited by 58 publications

(32 citation statements)

References 42 publications

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“…Our assay appeared to be equally as sensitive as the qPCR assay developed by Vaitomaa et al [35] for the detection of microcystins genes and to the assay developed by Rasmussen et al [41] for the detection of C. raciborskii. It is also one order of magnitude less sensitive than the assay developed by Al-Tebrineh et al [39] and Koskenniemi et al [46] for the detection of hepatotoxin-producing species.…”

Section: Discussionmentioning

confidence: 86%

“…A number of studies have developed Real Time PCR approaches for the detection and quantification of some of the genes belonging to the hepatotoxin synthetase clusters (mcy, nda and cyr) using SYBER-Green I and TaqMan technologies [38][39][40]. It was also demonstrated that the cylindrospermopsin synthetase gene C (cyrC) is useful for detecting and quantifying the toxic C. raciborskii in water [41,42].…”

Section: Discussionmentioning

confidence: 99%

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First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium <i>Aphanizomenon ovalisporum</i> in Water

Campo¹,

Lezcano²,

Agha³

et al. 2013

AiM

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Cylindrospermopsin (CYN) is an alkaloid that causes hepatotoxicity, neurotoxicity and general cytotoxicity in vertebrates. It is currently gaining widespread attention after its reported appearance in water bodies around the world. A. ovalisporum is capable of CYN-production and can form toxic blooms when favorable environmental conditions are available. We have developed for the first time a two-step qPCR assay using Taqman probes to detect and quantify potential CYN-producing A. ovalisporum in water samples. The assay was sensitive enough to discriminate between CYN-producing and non-CYN-producing A. ovalisporum in a mixed background, and discriminate between A. ovalisporum and other nostocales as C. raciborskii and A. bergii. The detection limit of the assay falls in the log linear range of 10 2 and 10 5 gene copies per reaction and is thus within the sensitivity range of previously published assays for the detection of other toxic cyanobacteria species. Our assay allows for the first time to quickly assess water quality for the presence of potentially CYN-producing A. ovalisporum and can be easily used for the purposes of monitoring water bodies.

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Section: Discussionmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 99%

First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium <i>Aphanizomenon ovalisporum</i> in Water

Campo¹,

Lezcano²,

Agha³

et al. 2013

AiM

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show abstract

“…For the molecular identification of the cyanobacteria, 16S rDNA oligonucleotide primers, CYA106F-CYA781R (Nubel et al, 1997) and ITS-23S rDNA P322-P340, (Iteman et al, 2000) were used and synthesized by (Bioneer, Korea). PCR reactions were performed as previously described for each primer pair (Al-Tebrineh et al, 2011;Iteman et al, 2000;Jungblut et al, 2006;Nubel et al, 1997).…”

Section: Identification and Amplification Of 16s Rdna And Its-23s Rdnmentioning

confidence: 99%

Characterization of blue green algae isolated from Egyptian rice field with potential anti-hepatitis C active components

Amer¹,

Wahab²,

Fathy³

et al. 2014

Afr. J. Biotechnol.

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Several species of cyanobacteria has been recognized for its therapeutic value that can be used for treatment of malnutrition, cancer and viral infection. Many natural occurring cyanobacteria are known to produce toxins, for example, species of the genera Microcystis, Nodularia, Nostoc, Anabaena, Aphanizomenon, Cylindrospermopsis, and Planktothrix (Oscillatoria). Cyanotoxins are classified according to their mode of action in vertebrates as hepatotoxins, neurotoxins, cytotoxins, dermatotoxins, and irritants. Microcystin is a hepatotoxin which commonly found in Microcystis and it was found to be produced by other genera, including Anabaena, Nostoc, Nodularia, and Planktothrix. In the present study cyanobacteria strain isolated from Egyptian soil was purified, characterized and identified as Nostoc sp. and named Nostoc EGY. PCR-based techniques targeting the toxin biosynthesis genes were used verifying absence of toxic genes in the newly purified cyanobacteria. Cell lysate was prepared from the purified strain; the efficacy of this lysate to prevent hepatitis C virus (HCV) replication in vitro was proved qualitatively and quantitatively. Lysate prepared from isolated cyanobacteria after 10 and 25 days of cultivation was able to prevent replication of in vitro cultivated HCV.

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“…To overcome this difficulty and better control the quality of PCR assay, an internal control has to be integrated into the rt-QPCR assay. Most of studies on cyanobacteria used 16S rRNA as an internal control to monitor PCR inhibition [1,17,25,32]. However, the wide range of variations of 16S rRNA level derived from total cyanobacteria made it difficult to assess the end-point of PCR results, suggesting that 16S rRNA may not be an appropriate quality control (QC) for detection of inhibitory effect in lake-water samples.…”

Section: Mcye Copy Numbers/mlmentioning

confidence: 99%

Rapid Detection and Quantitation of Microcystin-Producing Microcystis Using Real-Time PCR

Qiu¹,

Yuan²,

Kon³

et al. 2013

J Mol Biomark Diagn

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This study is to develop and validate a real-time quantitative PCR (rt-QPCR) assay for rapid quantitation of microcystin-producing Microcystis using unprocessed surface water samples collected from Alberta lakes. Microcystin synthetase gene E (mcyE) was targeted for microcystin-producing Microcystis and 16S rRNA was used to measure blooming level of total cyanobacteria. The assay was optimized and validated with 20 reference samples collected in 2011. The limit of detection (LOD) of rt-QPCR was 50 copies/ml for both mcyE and 16S rRNA. An excellent precision was observed in 24 replicates [coefficient of variation (cv)=1.12% for 1.0E+05 and = 0.79% for 1.0E+03 copies]. The rt-QPCR assay was applied for detection of mcyE in 527 water samples collected from 45 lakes during the open-water season of 2012 in Alberta and 369 samples were mcyE positive. Microcystin-producing Microcystis was detected in 41 out of the 45 lakes in which, relatively high copy numbers of mcyE (≥ 1.0E+05 copies/ ml) were determined in 9 lakes. Cyanobacteria were present in all 45 lakes determined by 16S rRNA. The rt-QPCR assay developed with specific target to mcyE is sensitive, specific and robust for rapid detection and differentiation of toxic Microcystis from non-toxic cyanobacteria in surface water.

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A new quantitative PCR assay for the detection of hepatotoxigenic cyanobacteria

Cited by 58 publications

References 42 publications

First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium <i>Aphanizomenon ovalisporum</i> in Water

First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium <i>Aphanizomenon ovalisporum</i> in Water

Characterization of blue green algae isolated from Egyptian rice field with potential anti-hepatitis C active components

Rapid Detection and Quantitation of Microcystin-Producing Microcystis Using Real-Time PCR

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A new quantitative PCR assay for the detection of hepatotoxigenic cyanobacteria

Cited by 58 publications

References 42 publications

First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium &lt;i&gt;Aphanizomenon ovalisporum&lt;/i&gt; in Water

First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium &lt;i&gt;Aphanizomenon ovalisporum&lt;/i&gt; in Water

Characterization of blue green algae isolated from Egyptian rice field with potential anti-hepatitis C active components

Rapid Detection and Quantitation of Microcystin-Producing Microcystis Using Real-Time PCR

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Product

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First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium <i>Aphanizomenon ovalisporum</i> in Water

First TaqMan Assay to Identify and Quantify the Cylindrospermopsin-Producing Cyanobacterium <i>Aphanizomenon ovalisporum</i> in Water