RRC ID 20873
Author Tie J, Uchigasaki S, Haseba T, Ohno Y, Isahai I, Oshida S.
Title Direct and rapid PCR amplification using digested tissues for the diagnosis of drowning.
Journal Electrophoresis
Abstract We developed a direct and rapid method for the diagnosis of death by drowning by PCR amplification of phytoplankton DNA using human tissues. The primers were designed based on the DNA sequence of the 16S ribosomal RNA gene (16S rDNA) of Cyanobacterium. Samples of lung, liver and kidney tissues were collected from 53 autopsied individuals diagnosed as death by drowning. Without DNA extraction, the tissue fragments were incubated directly in a digest buffer developed in this study, for 20 min. Using 1 microL of the tissue digest solution in PCR, the 16S rDNA was successfully amplified. The specific 16S rDNA fragment was identified from the standard picoplankton Euglena gracilis, the tissues of bodies died from drowning and water samples from the drowning scenes. On the other hand, no PCR products were found in the tissues of individuals who died from causes other than drowning. Various quantities of tissue weighing 1, 5, 10, 20 and 30 mg were tested, and the PCR amplification detected the specific 16S rDNA fragment from all the quantities of tissue tested. This method was found to be more reliable, sensitive, specific and rapid when compared to the conventional diagnosis of death by drowning using the diatom test by acid digestion method.
Volume 31(14)
Pages 2411-5
Published 2010-7
DOI 10.1002/elps.200900754
PMID 20564264
MeSH Base Sequence Cyanobacteria / genetics* DNA Primers* / genetics DNA, Bacterial / genetics DNA, Bacterial / isolation & purification Drowning / diagnosis* Euglena gracilis / genetics Euglena gracilis / isolation & purification Humans Kidney / microbiology Liver / microbiology Lung / microbiology Phytoplankton / genetics Phytoplankton / isolation & purification* Polymerase Chain Reaction / economics Polymerase Chain Reaction / methods* RNA, Ribosomal, 16S / genetics* Time Factors
IF 2.569
Times Cited 3
WOS Category CHEMISTRY, ANALYTICAL BIOCHEMICAL RESEARCH METHODS
Resource
Algae NIES-47