Background:rnGram stains are initially used as a pre-analytical indicator of specimen quality andrnacceptability for culture. It also gives the clinician preliminary information regarding the nature ofrnpotential pathogens present in thepatient specimen and thus serve to guide empirical therapy. Thisrnprocedure is still considered a high-complexity procedure by the Clinical Laboratory ImprovementrnAmendments (CLIA) program. The manual nature of the staining process and the subjectivity ofrnGram stain interpretation contribute to the incidence of errors. rnObjective:rnAssessment of Gram stain competency and associated factors among laboratoryrnprofessional in clinical laboratories at Mekelle city health facilityrn rnMethodology:rnA cross-sectional study was conducted from January 2020 to April 2020 on 77rndifferent health facilities present at Mekelle city. Convenient sampling method was used torncollectData from 148participants; of those are 81, 62 and 5 from private, governmental and uniformedrnhealth facility respectively.Qualitative data were collected from medical laboratory professional rnthrough a structured self-administered questionnaire and panel slide at on-site assessment of theirrnperformance. Question and panel slide was used to assess knowledge and skill of laboratoryrnprofessionals respectively. Panel slides were prepared from American Type CulturernCollection(ATCC)by using known bacterial strains and no organism slides were prepared from patientrnsample.Bloomâ€™s cut off point of59 % or below as low, 60-80 % as medium and above 80: as high wasrnused to describe the knowledge and practical skills of the respondents. rnResult:rnAmong 155 eligible medical laboratory professionals, 148 (95.5%) of them participatedandrnmost of them were males 78(52.7%). Onehundred twenty seven 127(85.2%), Nineteen 19(12.8%) andrn2(1.3%) participants scored low, medium and high skill level respectively. The level of skills andrnknowledge of medical laboratory professional had significant associated witheducational level,rnSLIPTA levelof health facility,training, higher institution type and sex had significant associationrnwithstudy participant about gram stain (P < 0.05). rnConclusion:rnThe present study showed that the majority of medical laboratory professional had lowrnknowledge and skill in gram stain examinations. Attention should be given to develop trainingrnstrategies that can improve laboratory professional knowledge and skill level. This could be achievedrnthrough pre service and in service training and also giving adequate emphasis to gram stain relatedrnpractical training, continuous followupand Regular competence assessment (supervision).