DNA Polymerase Technology (United States)

The objective of our participatory case study of community health worker (CHW) integration was to describe the role of CHWs in two local health departments (LHDs). Study partners co-developed an interview guide based on an existing framework. Fourteen staff participated in interviews. CHWs build community trust for LHDs, provide LHD core services, identify community concerns and priorities, and address social determinants of health. Integration barriers include hiring protocols, lack of understanding of the CHW role, inadequate compensation, and sustainable funding. The study identified key factors for an integration process that promotes the effectiveness, wellbeing, and sustainability of the workforce.

Potent PCR inhibitors in blood and soil samples can cause false negative results from PCR-based clinical and forensic tests. We show that the effect of these inhibitors is primarily upon Taq DNA polymerase, since mutational alteration of the polymerase can overcome the inhibition to the extent that no DNA purification is now required. An N-terminal deletion (Klentaq1) is some 10–100-fold inhibition resistant to whole blood compared to full-length, wild-type (w.t.) Taq, which is strongly inhibited by 0.1–1% blood. Further mutations at codon 708, both in Klentaq 1 and Taq, confer enhanced resistance to various inhibitors of PCR reactions, including whole blood, plasma, hemoglobin, lactoferrin, serum IgG, soil extracts and humic acid, as well as high concentrations of intercalating dyes. Blood PCR inhibitors can predominantly reduce the DNA extension speed of the w.t. Taq polymerase as compared to the mutant enzymes. Single-copy human genomic targets are readily amplified from whole blood or crude soil extract, without pretreatment to purify the template DNA, and the allowed increase in dye concentration overcomes fluorescence background and quenching in real-time PCR of blood.