Extraction of Human Nuclear DNA from Feces Samples Using the Qiaamp DNA Stool Mini Kit

Vandenberg, Nicholas; Oorschot, Roland A.H. van

doi:10.1520/jfs15502j

Cited by 18 publications

(9 citation statements)

References 9 publications

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“…It can be difficult to purify DNA from stool samples due to hardness of the cyst wall and the presence of inhibiting substances, which can inhibit the Taq polymerase. 13,[30][31][32] According to Campos-Górgora et al, 33 the major component of the Entamoeba cyst wall is chitin, a homopolymer of beta-(1,4)-linked N-acetyl-D-glucosamine that confers great rigidity and resistance. Therefore, the utilization of both chemistry and physical conditions was sufficient to yield a great quantity of genetic material to proceed with the PCR.…”

Section: Discussionmentioning

confidence: 99%

Validation and utilization of PCR for differential diagnosis and prevalence determination of Entamoeba histolytica/Entamoeba dispar in Salvador City, Brazil

Santos¹,

Gonçalves²,

Soares³

2011

Braz J Infect Dis

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Section: Discussionmentioning

confidence: 99%

Validation and utilization of PCR for differential diagnosis and prevalence determination of Entamoeba histolytica/Entamoeba dispar in Salvador City, Brazil

Santos¹,

Gonçalves²,

Soares³

2011

Braz J Infect Dis

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“…14,19,20 These kits use a lysis buffer which contains a detergent to disrupt cellular membranes, and a protease (proteinase K) for digestion of cellular and intracellular proteins. Digestion with lysozyme was also used for the breakdown of the Gram-positive bacteria cell walls.…”

Section: Dna Extraction Proceduresmentioning

confidence: 99%

Lyophilisation improves the extraction of PCR‐quality community DNA from pig faecal samples

Ruíz

Rubio

2009

J Sci Food Agric

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BACKGROUND: Faeces are increasingly used as sources of DNA for genetic and ecological studies. Although multiple methods to preserve faecal samples prior to DNA extraction have been used (e.g., 70% or absolute ethanol, freezing at −20• C or in liquid nitrogen) no information is at present available in the literature on the use of lyophilised faeces. Accordingly, the yield and quality of the community DNA obtained by using four different commercial DNA extraction kits (QIAamp DNA Stool Mini Kit, REALPURE Spin Kit, SPEEDTOOLS Tissue DNA Extraction Kit, and JETQUICK Tissue DNA Spin Kit) from fresh and lyophilised samples of faeces were studied here.

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“…Total nucleic acid was extracted from the stool specimens using QIAamp DNA Stool Mini Kit (QIAGEN, Germany) according to the manufacturer's instructions [18] and subsequently stored at -20°C. The extracted stool DNA was then shipped to the University of Calgary, in Calgary, Alberta, for molecular analysis using the Allplex™…”

Section: Molecular Testingmentioning

confidence: 99%

Molecular detection of Enteropathogens from diarrheic stool of HIV positive patients in Gondar, Ethiopia

et al. 2018

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BackgroundInfectious diarrhea is a common problem in the developing world, especially among people living with HIV/AIDS. Traditional diagnostic methods such as stool culture and microscopic examination are limited by resources and poor sensitivity. The use of molecular diagnostics for enteropathogen detection in this region of sub-Saharan Africa has not been fully explored. We sought to identify risk factors and characterize enteropathogens from diarrheic stools of HIV-positive patients in Gondar, Ethiopia using multiplex molecular panels targeting key infectious agents.MethodsA cross-sectional study of 100 stool samples was performed. Samples were collected consecutively from HIV- positive patients presenting with diarrhea at University of Gondar Hospital clinic, a major center in NW Ethiopia. Genomic DNA was extracted from stool and processed using a multiplex molecular panel Allplex™ [Seegene, Canada]. Correlations between patient characteristics, symptoms, public health risk factors, and enteropathogen type (s) were studied. Eighty-six samples were successfully analyzed by molecular methods.ResultsThe mean age was 35 with 43% male. Eighty percent lived in an urban area, 18% had access to well water only, and 81% practiced proper hand hygiene. The majority of patients (72%) were receiving HAART with a median CD4 cell count of 362/μL. Multiple pathogens were detected in 94% of specimens, with an average of 5 enteropathogens per sample. Common bacteria, viruses, and parasites detected were Shigella spp./enteroinvasive E. coli (80%), enterotoxigenic E. coli (73%), Norovirus (16%) and B. hominis (62%). CD4 cell count < 500/ μL was associated with the presence of viruses (p = 0.004) and the absence of STEC (p = 0.010). The use of HAART or CD4 levels was not associated with the number of enteropathogens detected.ConclusionsDiarrheic stool from HIV-positive outpatients in Gondar, Ethiopia had on average 5 enteropathogens present in their stool. Shigellaspp./enteroinvasive E. coli and enterotoxigenic E. coli are the major pathogens, not dissimilar to immunocompetent individuals in low income countries.

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Extraction of Human Nuclear DNA from Feces Samples Using the Qiaamp DNA Stool Mini Kit

Cited by 18 publications

References 9 publications

Validation and utilization of PCR for differential diagnosis and prevalence determination of Entamoeba histolytica/Entamoeba dispar in Salvador City, Brazil

Validation and utilization of PCR for differential diagnosis and prevalence determination of Entamoeba histolytica/Entamoeba dispar in Salvador City, Brazil

Lyophilisation improves the extraction of PCR‐quality community DNA from pig faecal samples

Molecular detection of Enteropathogens from diarrheic stool of HIV positive patients in Gondar, Ethiopia

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