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A total of 9 populations of tropical bed bug (Cimex hemipterus) collected from urabn area in Penang and other locations in Malaysia. The populations involved in this study included Sungai Ara, Taman Brown, Desa Permai Indah, Tasek Gelugor and Lahar Tembun in Penang, Shah Alam, Petaling Jaya, Pahang and Seremban, Malaysia. Deoxyribonucleic Acid (DNA) was extracted from total 5 individual bed bugs each location and used as a template for the Polymerase Chain Reaction (PCR) to amplify 5 different genomic microsatellite loci. PCR products were resolved by agarose gel electrophoresis and visualized with ethidium bromide staining. The genotyping data from the sample sites revealed that PCR-based genotyping reliably separated the samples into genotypic groups corresponded to the location.