Title |
The Gut Microbiotassay: a high-throughput qPCR approach combinable with next generation sequencing to study gut microbial diversity
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Published in |
BMC Genomics, November 2013
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DOI | 10.1186/1471-2164-14-788 |
Pubmed ID | |
Authors |
Marie Louise Hermann-Bank, Kerstin Skovgaard, Anders Stockmarr, Niels Larsen, Lars Mølbak |
Abstract |
The intestinal microbiota is a complex and diverse ecosystem that plays a significant role in maintaining the health and well-being of the mammalian host. During the last decade focus has increased on the importance of intestinal bacteria. Several molecular methods can be applied to describe the composition of the microbiota. This study used a new approach, the Gut Microbiotassay: an assembly of 24 primer sets targeting the main phyla and taxonomically related subgroups of the intestinal microbiota, to be used with the high-throughput qPCR chip 'Access Array 48.48', AA48.48, (Fluidigm®) followed by next generation sequencing. Primers were designed if necessary and all primer sets were screened against DNA extracted from pure cultures of 15 representative bacterial species. Subsequently the setup was tested on DNA extracted from small and large intestinal content from piglets with and without diarrhoea. The PCR amplicons from the 2304 reaction chambers were harvested from the AA48.48, purified, and sequenced using 454-technology. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Netherlands | 1 | 14% |
Germany | 1 | 14% |
Ireland | 1 | 14% |
India | 1 | 14% |
Unknown | 3 | 43% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 5 | 71% |
Practitioners (doctors, other healthcare professionals) | 2 | 29% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 5 | 2% |
Germany | 3 | 1% |
Denmark | 2 | <1% |
India | 1 | <1% |
Japan | 1 | <1% |
Sweden | 1 | <1% |
Unknown | 225 | 95% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 46 | 19% |
Student > Master | 46 | 19% |
Student > Ph. D. Student | 40 | 17% |
Student > Bachelor | 16 | 7% |
Student > Postgraduate | 11 | 5% |
Other | 38 | 16% |
Unknown | 41 | 17% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 99 | 42% |
Biochemistry, Genetics and Molecular Biology | 32 | 13% |
Immunology and Microbiology | 17 | 7% |
Medicine and Dentistry | 14 | 6% |
Veterinary Science and Veterinary Medicine | 5 | 2% |
Other | 19 | 8% |
Unknown | 52 | 22% |