Loop Mediated Isothermal PCR (LAMP) for the Detection of Borrelia from Blood Cultures

Abstract

Borrelia organisms were harvested from blood culture and tested using single primers and loop mediated isothermal PCR (LAMP). Samples were harvested at either 6 days or at 8 weeks and DNA was extracted using one of three methods. In addition, pre-processing steps included Liberase treatments to release organisms from any residual attached collagen and human DNA removal. Combinations of processes were used to determine the best procedures for harvesting organisms, processing the pellet, and extracting DNA prior to using PCR. Of the total number of positive signals obtained, 9 of 45 (20%) gave positive signals when using plasmid primers specific for B. burgdorferi, 14 of 40 (35%) were positive when using OspC primers, and 25 of 47 (53%) were positive when using LAMP primers for the DNA that codes for 16S rRNA.