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STR-Typing of Human DNA from Human Fecal Matter Using the QIAGEN QIAamp

STR-Typing of Human DNA from Human Fecal Matter Using the QIAGEN QIAamp,Donald J. Johnson,Liane R. Martin,Katherine A. Roberts

STR-Typing of Human DNA from Human Fecal Matter Using the QIAGEN QIAamp  
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The purpose of this study was to compare the effectiveness of the QIAGEN QIAamp R � Stool Mini Kit against a standard phenol- chloroform procedure for the extraction, quantitation, and STR-typing of human nuclear DNA from human feces. Stools from six subjects were sampled by swabbing and excision. Samples extracted with the QIAamp kit gave a wide range of DNA yields, whereas those extracted by the organic method yielded no DNA. DNA was not recovered from one subject's stools by either procedure. The QIAamp extracts were amplified with the Profiler PlusTM and COfilerTM kits, and PCR inhibition was observed with DNA extracts that were further concentrated. Substitution of water or TE-4 for the QIAamp elution buffer eliminated most, if not all, of the inhibition. A modified QIAamp procedure was used to extract thirty samples, which were subjected to one of five environmental conditions. DNA was recovered from all of these samples, and typing results were obtained on 93% of the samples. Fecal matter is a less considered but potentially significant item of evidence. It is encountered in various casework situations, from trace quantities to entire stool deposits. In sexual assault cases, for example, small quantities of fecal material can be transferred to the sodomite's penis or other objects inserted anally. Contrastingly, a criminal may intentionally or unintentionally defecate at a crime scene, to leave an entire bowel movement. In many cases, the identification and individualization of fecal matter can establish the link between the victim and assailant. Fecal matter is the end product of digestion. Digestion and ab- sorption are functions of the gastrointestinal (GI) tract, which is basically a muscular tube that extends from the mouth to the anus. The GI tract is lined with epithelium (cell layer) and covered with peritoneum (membrane) along the majority of its length. The epithe- lial lining is regenerated every two to six days (4), and an estimated 17 billion cells are shed per day by the small intestine in humans (1). The intestinal lining is comprised of two distinct epithelial cell types: the columnar absorptive cell and the goblet cell. Defoliated cells that are not destroyed by the digestive process are excreted intact, although their morphology can be significantly altered (5). In addition, nucleated squamous epithelial cells can be transferred from the lining of the anal canal to the passing stool during the process of defecation (5). Feces produced from an average diet is approximately 75% wa- ter and 25% solid material. The solid component includes bacte- ria (∼30%); inorganic material, mainly calcium and phosphates
Published in 2005.
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