Abstract: The present study was performed to investigate the phenotypic resistance of all isolated species of Staphylococcus to macrolide-lincosamide streptogramin-B through double disc test. One hundred eighty one strains of staphylococci, consisting 59 Staphylococcus aureus and 122 Coagulase-Negative Staphylococci (CNS) were collected between March and April 2013 from environmental sources such as gymnasia, shopping malls and ATM key in the Qasssim region and study was conducted at College of Applied Medical Sciences, Qassim University, Saudi Arabia. All isolates were subjected to in vitro anti-microbial susceptibility testing by using Kirby Bauer Method as per Clinical and Laboratory Standards Institute guidelines. The prevalence rate of total MLS resistance phenotype was observed in 29.8% of Methicillin-Resistant (MR) and Methicillin-Sensitive (MS) Staphylococcus species. The constitutive (cMLS_B ), inducible (iMLS_B ) and MS phenotypes prevalence was in 16.9, 16.9 and 22%, respectively among Staphylococcus aureus. While prevalence of constitutive (cMLS_B ), inducible (iMLS_B ) and MS phenotypes in coagulase negative Staphylococcus was 2.4, 3.2 and 16.3%, respectively. The high prevalence of cMLS_B and inducible Clindamycin iMLS_B resistance phenotypes in Methicillin resistance staphylococci as compared to Methicillin sensitive isolates was noticed in Qassim Province of Saudi Arabia. Therefore, we recommend that routine screening of staphylococci isolates by double-disc test is critical and necessary before the treatment so that therapeutic failure may be avoided.