Coprocytobiology: A Technical Review of Cytological Colorectal Cancer Screening in Fecal Samples

Ryan, Liam; Wong, Yen Ting; Dwyer, Karen M.; Clarke, Deborah; Kyprian, Lynda; Craig, Jeffrey M.

doi:10.1177/24726303211024562

Cited by 6 publications

(6 citation statements)

References 77 publications

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“…Instead, we hypothesise that the target animal cells and therefore their DNA would be heterogeneously distributed throughout the faecal mass. Since faeces are unformed in the upper regions of the colon, exfoliated epithelial cells from those regions would likely be incorporated within the faecal mass through peristaltic contractions of the colon [ 13 , 66 ].…”

Section: Discussionmentioning

confidence: 99%

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A new DNA extraction method (HV-CTAB-PCI) for amplification of nuclear markers from open ocean-retrieved faeces of an herbivorous marine mammal, the dugong

Ooi¹,

McMichael²,

Hunter³

et al. 2023

PLoS ONE

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Non-invasively collected faecal samples are an alternative source of DNA to tissue samples, that may be used in genetic studies of wildlife when direct sampling of animals is difficult. Although several faecal DNA extraction methods exist, their efficacy varies between species. Previous attempts to amplify mitochondrial DNA (mtDNA) markers from faeces of wild dugongs (Dugong dugon) have met with limited success and nuclear markers (microsatellites) have been unsuccessful. This study aimed to establish a tool for sampling both mtDNA and nuclear DNA (nDNA) from dugong faeces by modifying approaches used in studies of other large herbivores. First, a streamlined, cost-effective DNA extraction method that enabled the amplification of both mitochondrial and nuclear markers from large quantities of dugong faeces was developed. Faecal DNA extracted using a new ‘High Volume- Cetyltrimethyl Ammonium Bromide- Phenol-Chloroform-Isoamyl Alcohol’ (HV-CTAB-PCI) method was found to achieve comparable amplification results to extraction of DNA from dugong skin. As most prevailing practices advocate sampling from the outer surface of a stool to maximise capture of sloughed intestinal cells, this study compared amplification success of mtDNA between the outer and inner layers of faeces, but no difference in amplification was found. Assessment of the impacts of faecal age or degradation on extraction, however, demonstrated that fresher faeces with shorter duration of environmental (seawater) exposure amplified both markers better than eroded scats. Using the HV-CTAB-PCI method, nuclear markers were successfully amplified for the first time from dugong faeces. The successful amplification of single nucleotide polymorphism (SNP) markers represents a proof-of-concept showing that DNA from dugong faeces can potentially be utilised in population genetic studies. This novel DNA extraction protocol offers a new tool that will facilitate genetic studies of dugongs and other large and cryptic marine herbivores in remote locations.

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Section: Discussionmentioning

confidence: 99%

“…The bulk of mammalian faeces comprises water and soluble molecules such as mucin, and the remainder consists of undigested food and microflora [ 13 ]. A small portion of faeces is composed of epithelial cells sloughed from the host’s gastrointestinal tract [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

A new DNA extraction method (HV-CTAB-PCI) for amplification of nuclear markers from open ocean-retrieved faeces of an herbivorous marine mammal, the dugong

Ooi¹,

McMichael²,

Hunter³

et al. 2023

PLoS ONE

View full text Add to dashboard Cite

show abstract

Section: Discussionmentioning

confidence: 99%

“…The bulk of mammalian faeces comprises water and soluble molecules such as mucin, and the remainder consists of undigested food and microflora [12]. A small portion of faeces comprises epithelial cells sloughed from the host’s gastrointestinal tract [13].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Development of a new faecal DNA extraction method (HV-CTAB-PCI) for amplification of mitochondrial and nuclear markers used in genetic analyses of dugongs (Dugong dugon)

Ooi

McMichael

Hunter

et al. 2022

Preprint

View full text Add to dashboard Cite

Non-invasively collected faecal samples are an alternative source of DNA to tissues, that may be used in genetic studies of wildlife when direct sampling of animals is difficult. Although several faecal DNA extraction methods exist, their efficacy varies between species. Previous attempts to amplify mitochondrial DNA (mtDNA) markers from faeces of wild dugongs have met with limited success and nuclear markers (microsatellites) have been unsuccessful. This study aimed to develop a new tool for sampling both mtDNA and nuclear DNA (nDNA) from dugong faeces by modifying approaches used in studies of other large herbivores. First, amplification success of genetic markers from dugong faeces was compared between an established QIAamp and a newly developed DNA extraction method. Faecal DNA extracted using a new 'High Volume-CTAB-PCI' (HV-CTAB-PCI) method was found to achieve comparable amplification results to extraction of dugong skin DNA. As most prevailing practices advocate sampling from the outer surface of a stool to maximise capture of sloughed intestinal cells, this study compared amplification success of mtDNA between the outer and inner layers of faeces, but no difference in amplification was found. Assessment of the impacts of faecal age or degradation on extraction, however, demonstrated that fresher faeces with shorter duration of environmental (seawater) exposure amplified mtDNA and nDNA better than eroded scats. Using the HV-CTAB-PCI method, nDNA was successfully amplified for the first time from dugong faeces. This novel DNA extraction protocol offers a new tool that will facilitate genetic studies of dugongs and other large and cryptic marine herbivores in remote locations.

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Effectiveness of Fecal DNA Syndecan-2 Methylation Test in Detecting Colorectal Cancer in High-Risk Population in China

Luo,

Lin,

Jiao

et al. 2023

Preprint

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Purpose:To validate the effectiveness of fecal DNA Syndecan-2 methylation (mSDC2) test in the detection of colorectal cancer in high-risk population in China. This may provide evidence-based data of diagnostic or screening for colorectal cancer in China. Methods A high-risk Chinese pilot comprising 1130 individuals aged 40-79 was registered for colorectal cancer (CRC) detection using the fecal mSDC2 test. The sensitivity and specificity to CRC, advanced adenomas (AA), and advanced colorectal neoplasia (ACN) were estimated. High-risk factors for incidence of colorectal lesions were analyzed and then were constructed a logistic regression model to better reflect the efficacy of the fecal mSDC2 test. ResultsAccording to established criteria, 1035 high-risk individuals were included for analysis. Among them, 16 CRCs (1.55%), 65 AAs (6.28%), 189 NAAs (18.26%), and 150 polyps (14.49%) were defined by colonoscopy and pathological examination. The sensitivity of mSDC2 test to CRC and AA was 87.50% and 40.00%. The specificities were 95.61% in the others group. The positive predictive value (PPV) of mSDC2 test for CRC, AA, and ACN was 16.09%, 29.89% and 45.98%, respectively. In addition, the negative predictive value (NPV) of mSDC2test for CRC was 99.79%. The positive of mSDC2 test is a significant risk signal for the occurrence of ACN (P <0.001) after adjusting other high-risk covariates. ConclusionThe results of this CRC screening showed that offering a combination of fecal mSDC2test and colonoscopy may be an active option to early detect malignant lesions of the colorectum in Chinese high-risk population.

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Coprocytobiology: A Technical Review of Cytological Colorectal Cancer Screening in Fecal Samples

Cited by 6 publications

References 77 publications

A new DNA extraction method (HV-CTAB-PCI) for amplification of nuclear markers from open ocean-retrieved faeces of an herbivorous marine mammal, the dugong

A new DNA extraction method (HV-CTAB-PCI) for amplification of nuclear markers from open ocean-retrieved faeces of an herbivorous marine mammal, the dugong

Development of a new faecal DNA extraction method (HV-CTAB-PCI) for amplification of mitochondrial and nuclear markers used in genetic analyses of dugongs (Dugong dugon)

Effectiveness of Fecal DNA Syndecan-2 Methylation Test in Detecting Colorectal Cancer in High-Risk Population in China

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