Four out of six animals in group A became negative within 1 day post treatment with both ITS1 TD PCR and HCT (Table 1). After 1 day of treatment, one animal was positive in ITS1 TD PCR and negative in HCT, and another animal was positive in HCT and negative in ITS1 TD PCR. From 2 days post-treatment till the end see more of the sampling period (44 days after first treatment), all animals were negative in both the ITS1 TD PCR and the HCT. In group B, all six animals were occasionally positive in ITS1 TD PCR and/or HCT during the follow-up period between 1 and 16 days after treatment and even after retreatment (on day 19) up to day 44 after treatment (Table 2). The parasite
detection rate of ITS1 TD PCR after two treatments in animal group B was higher than that of HCT. Positivity rate of ITS1 TD PCR was 84.85% (56/66), while HCT was 57.58% (38/66). The ITS1 TD PCR could detect relapse up to three days earlier than microscopical parasite detection. Livestock production is considered Autophagy inhibitor the main lifeline for millions of families in many sub-Saharan countries. However, the emergence of drug resistant trypanosomes
presents a serious threat to agriculture in the regions. Therefore, studies on drug resistance and development of novel compounds against trypanosomes are necessary for effective control. These studies greatly benefit from rapid and cost-efficient molecular tools to detect the presence of trypanosomes. Compared to microscopy, PCR-based assays have the advantage that they are more amenable to high throughput processing and that specimens can be stored longer term. In addition, differentiation between trypanosome taxa by microscopy is much more cumbersome than with molecular methods, such as PCR. This study presents an improved ITS1-based
PCR assay for diagnosis of trypanosomosis and for efficacy assessment of trypanocidal compounds where the detection of genomic rDNA of Trypanosoma specific ITS1 serves as surrogate for parasite detection but with higher L-NAME HCl analytical sensitivity. The same primer sequences were employed in a previous survey on AAT in Ethiopia ( Fikru et al., 2012). However, the reaction mixture and cycling conditions of the ITS1 TD PCR are refined for optimal sensitivity and specificity. The “Touchdown” approach that employs more stringent primer-template hybridisation conditions is introduced to minimise potential non-specific amplifications. It favours amplification of desirable products during early cycles that will out-compete potential non-specific products during the remaining cycles ( Korbie and Mattick, 2008). For T. congolense, the newly developed ITS1 TD PCR has a lower detection limit of 10 parasites/ml blood while for T. vivax, this is 100 parasites/ml blood. A similar difference in detection limits between T. congolense and T.