Using species-specific repeat and PCR–RFLP in typing of DNA derived from blood of human and animal species

Abstract

Species determination of tissue specimens, including blood, is an important component of forensic analysis to distinguish human from animal remains. DNA markers based on a method of species-specific PCR and amplifying the 359-base pair (bp) fragment of the mitochondrially encoded cytochrome-b gene and then digestion with the TaqI restriction enzyme were developed for detection and discrimination of human, cattle, buffalo, horse, sheep, pig, dog, cat and chicken blood samples. The results reveal that PCR-amplification of the gene encoding the species-specific repeat (SSR) region generated 603 bp in cattle and buffalo, 221 bp in horse, 374 bp in sheep, <or=100 bp in pig, 808 bp in dog, 672 bp in cat and 50 bp in chicken. Restriction analysis of the amplified 359-bp portion of the cytochrome-b gene using the TaqI restriction enzyme results in species-specific restriction fragment length polymorphism (RFLP) between buffalo, cattle and human. Two different bands were generated in buffalo (191 and 168 bp) and human (209 and 150 bp), with no digestion in cattle (359 bp). Cytochrome-b is a highly conserved region and consequently a good molecular marker for diagnostic studies. Therefore, the two complementary techniques, SSR-PCR and PCR-RFLP, could be used successfully as routine methods in forensics for sensitive, rapid, simple and inexpensive identification of the species in bloodstains.

Neuigkeiten zur praktischen ABC-Blutgruppen-Bestimmung bei Katzen

“…While immunological typing is generally sufficient when typing recipient and donor cats in practice, genotyping is preferred or performed in combination with immunological typing to assure the detection of the recessive alleles b and a c in type A and C cats when breeding cats [7]. Similarly, regular blood typing methods are complemented by genotyping assays to assure accuracy in blood typing in humans for several blood groups [32] [33]. Finally, crossmatching may be recommended prior to transfusing untyped and even A-B matched cats due to the potential presence of other naturally occurring alloantibodies and particularly for repeated transfusions > 4 days after the first transfusion [12][13] [34].…”

Non-specific peaks generated by animal DNA during human STR analysis: Peak characteristics and a novel analysis method for mixed human/animal samples

“…In previous studies involving sequence analysis of 12S or 16S ribosomal RNA (rRNA) [2,3], methods using species-specific oligonucleotides (SSOs) [4] and classification by restriction fragment length polymorphisms (RFLPs) [5] were developed for identifying animal species in forensic settings. Currently, in wildlife forensic science, identification of animal species is commonly performed using cytochrome b (Cytb) or cytochrome c oxidase subunit I (COI) in mitochondrial DNA [6][7][8].…”

Identifying the species of origin in commercial sausages in South Korea

“…Recently, molecular methods have been developed for DNA extracted from various sources and used for many purposes (e.g. economic, social, religious reasons, scientific, or industrial) (Bataille et al 1999;Kusama et al 2004;Girish et al 2005;El-Sayed et al 2010;Ali et al 2012).…”