When
DNA extracts from bovine blood were added to the templates, both pCS20 and sodB LAMP could not detect 10 copies in two samples, which is in accordance with real-time PCR (Table 2). When DNA extracts from A. variegatum were added to the templates, both pCS20 and sodB LAMP failed in detecting 10 copies in all five samples, while real-time PCR could detect in four. The pCS20 PCR using the KAPA Blood PCR kit detected more positives than the pCS20 PCR using the AmpliTaq Gold PCR kit in the templates with 102 and 103 copies (Table 2). Table 2 Inhibitory effects of DNA
extracts from field Selleck IWR1 samples on pCS20 PCR, pCS20 real-time PCR, pCS20 LAMP, and sodB LAMP No. of samples: Sample type No. of plasmid Screening Library copies per reaction Tested pCS20 PCR positive pCS20 real-time PCR positive pCS20 LAMP positive sodB LAMP positive DNA extracts from bovine blood 1 5 0 (0)a 0 0 0 10 5 0 (0) 3 3 3 102 5 2 (0) 5 4 5 103 5 5 (0) 5 5 5 104 5 5 (5) 5 5 5 DNA extracts from Amblyomma variegatum 1 5 0 (0) 0 0 0 10 5 0 (0) 4 0 0 102 5 5 (0) 5 BGB324 price Rho 5 5 103 5 5 (3) 5 5 5 104 5 5 (5) 5 5 5 aTotal no. of samples positive
by using the KAPA Blood PCR kit (Total no. of samples positive by using the AmpliTaq Gold PCR kit). Detection of E. ruminantium DNA in field samples A total of 140 A. variegatum ticks were collected in 7 sites in Uganda and individually analyzed for the presence of E. ruminantium DNA. Out of 140 ticks, including 96 males and 44 females, 12 ticks (11 male and 1 female) were found positive with both pCS20 LAMP and sodB LAMP. The pCS20 real-time PCR detected 13 positives, including the 12 LAMP-positive ticks and an additional tick from Dokolo, while pCS20 PCR could detect only 8 positives (Table 3). All the samples found positive with PCR were also positive with LAMP. The percent positive with LAMP (8.57%) was higher than with PCR (5.71%) but slightly lower than with real-time PCR (9.29%). Of the 150 bovine, 35 goat, and 19 lamb blood samples analyzed, two lamb samples were positive using PCR, real-time PCR, and LAMP (Table 3). Table 3 Comparison of pCS20 PCR, pCS20 real-time PCR, pCS20 LAMP, and sodB LAMP for the detection of E. ruminantium in various field samples No.