Trypanosoma brucei s.l.: Characterisation of stocks from Central Africa by PCR analysis of mobile genetic elements

The typing of Trypanosoma evansi isolates using mobile genetic element (MGE) PCR

“…However, the appearance of hostdependent clusters (Fig. 2) indicates that RIME may not be a useful marker in determination of T. evansi genotypes and especially if the isolates have a different host origin, unlike with other members of subgenus Trypanozoon (Tilley et al 2003;Simo et al 2005). To check whether the observed positional variation affected the RIME targeting PCR (Tilley et al 2003) or loop-mediated isothermal amplification test (Njiru et al 2008), all the serially passaged isolates were analyzed (Table 1).…”

“…The MGE-PCR amplifies the element flanking regions through a combination of specific priming within the element, priming with adjacent elements, and mispriming in flanking regions (Hide and Tilley 2001). The technique has successfully been used to categorize the T. brucei isolates into three distinct groups reflecting human infectivity status and geographical isolation (Tilley et al 2003) and to reveal microgenetic variability between Trypanosoma brucei gambiense isolates from west Africa (Simo et al 2005). Despite the success achieved with MGE-PCR in genotyping some members of subgenus Trypanozoon namely T. brucei rhodesiense, T. brucei gambiense and T. brucei brucei, the test has not been used in the analysis of T. evansi isolates.…”

New developments in human African trypanosomiasis

“…Clearly, this mindset has not helped us gain a realistic view of sleeping sickness epidemiology in west Africa. Pigs have been shown repeatedly to harbour T. brucei gambiense [36 ]. Even though pigs can clear infections in less than 6 months, as Penchenier et al [43] demonstrate, they are still capable of maintaining parasite populations outside the human cycle.…”

“…Two new molecular markers promise to overcome these limitations and greatly improve our ability to study different T. brucei genotypes in the field. Simo et al [36 ] demonstrated the utility of mobile genetic element PCR (originally developed by Hide and Tilley [37]) to distinguish different strains of T. brucei across all three 'subspecies' and notably among T. brucei gambiense, which is at the same time the most medically relevant and the least genetically variable T. brucei subspecies. The beauty of this approach is that it requires only a single PCR, and is thus fast and relatively inexpensive.…”

Arboprotozoae

“…Appropriate analyses show that sub-populations of T. cruzi are present in roughly 12% of the cases, and provide proof for the major genetic heterogeneity of the circulating trypanosomes [75]. T. brucei: A PCR was described taking into account the heterogeneity of the Trypanosoma strains present in Cameroon, and amplifying sectors within the mobile genetic elements [76], furthermore, a real-time PCR in which the 177 bp repeat satellite DNA is detected [77]. The PCR analysis was able to prove that an infection with T. brucei was established in the tsetse fly after 11 days.…”