Research articles
ScienceAsia 52 (2026): 1-7 |doi:
10.2306/scienceasia1513-1874.2026.053
Confirmatory assay for sambar deer species identification by
pyrosequencing analysis of SNP markers on cytochrome b
gene
Natchaya Buakhaoa, Chavin Chaisongkramb, Khemika Lomthaisonga,*
ABSTRACT: Sambar deer (Rusa unicolor) is a protected species in Thailand. An assay based on PCR amplification of
cytochrome b (Cyt b) gene has recently been reported for screening to determine sambar deer origin from specimens.
As rusa deer (Rusa timorensis) and sika deer (Cervus Nippon) were also detected, species origin confirmation of sambar
deer needs to be further analyzed and refined. Thus, this study aims to differentiate sambar deer from rusa and sika
deer through SNPs analysis on partial Cyt b gene (366 bp) using pyrosequencing. The amplified Cyt b genes from all
three species were sequenced and compared to determine the informative SNP markers. The sequencing primer was
subsequently designed and used for species identification through pyrosequencing. The results showed that species
origin could be accurately determined via four bases. Remarkably, sambar deer origin specimens could be discriminated
from those of rusa deer and sika deer. Thepyrosequencingassayforspecies identification of sambar deer is reproducible
and robust for use with mixedmeat, cookedanduncookedmeatsamples. Inaddition, thesambardeeroriginofpositive
samples from screening assay could be indicated by this method, and consequently can be used as a confirmatory assay
for sambar deer origin in specimens.
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| a |
Forensic Science Program, Department of Integrated Science, Faculty of Science, Khon Kaen University,
Khon Kaen 40002 Thailand |
| b |
Khon Kaen Zoo, Zoological Park Organization, Khon Kaen 40000 Thailand |
* Corresponding author, E-mail: khemlo@kku.ac.th
Received 27 May 2025, Accepted 19 May 2026
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