Background: Isoniazid Preventive Therapy (IPT) reduced Tuberculosis (TB) among People Living with HIV (PLHIV). Despite this, uptake has been reported to be sub-optimal . We describe characteristics of visits in which PLHIV were screened TB negative (as the main source for IPT initiation), determine characteristics of visits in which PLHIV were initiated on IPT as well as determined factors associated with IPT initiation to inform program scale up and improve quality of service.Methods : Retrospective cohort study design which involved PLHIV enrolled into care and treatment clinics in Dar es Salaam, Iringa and Njombe regions from January 2012 to December 2016. The study aimed at evaluating implementation of IPT among PLHIV. Data analysis was conducted using STATA.Results: A total 173,746 were enrolled in CTC in the 3 regions during the period of follow up and made a total of 2,638,876 visits. Of the eligible visits, only 24,429 (1.26%) were initiated on IPT. In multivariate analysis, 50 years and more (aOR=3.42, 95% CI: 3.07-3.82, P<0.01), bedridden functional status individuals with bedridden functional status (aOR=4.56, 95% CI:2.45-8.49, P<0.01) and WHO clinical stage II had higher odds of IPT initiation (aOR=1.18, 95% CI:1.13-1.23, P<0.01). Furthermore, enrolment in 2016 (aOR=2.92, 95% CI:2.79-3.06, P<0.01), enrolment in hospitals (aOR=1.84, 95% CI:1.77-1.90, P<0.01), enrolment in public health facilities (aOR=1.82, 95% CI: 1.75-1.90, P<0.01) and been on care for more than one year (aOR=6.77, 95% CI: 5.25-8.73, P<0.000) were also more likely to be initiated on IPT. Enrollment in Iringa (aOR=0.44, 95% CI: 0.41-0.47, P<0.01) and good adherence (aOR=0.56, 95% CI 0.47-0.67, P<0.01) was less likely to be initiated on IPT.Conclusions: Our study documented low IPT initiation proportion among those who were enrolled in HIV care and eligible in the 3 regions during the study period. Variations in IPT initiation among regions signals different dynamics affecting IPT uptake in different regions and hence customized approaches in quality improvement. Implementation research is needed to understand health system as well as cultural barriers in the uptake of IPT intervention.