Application of PCR and DNA probes in the characterisation of trypanosomes in the blood of cattle in farms in Morogoro, Tanzania

Mugittu, Kefas; Silayo, R.S.; Majiwa, Phelix A.O.; Kimbita, EN; Mutayoba, Benezeth; Maselle, R.M.

doi:10.1016/s0304-4017(00)00365-4

Cited by 25 publications

(12 citation statements)

References 28 publications

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“…In our analysis, we picked out four isolates that had been previously categorised as single infections to be mixed infections. Direct sequencing of these products showed that KETRI 2729 contained a T. vivax [not the West African type because it was negative with TVW1 and TVW2, Masiga et al (1992) but positive with universal T. vivax primers (Masake et al 1997;Morlais et al 2001] (Mugittu et al 2000), while triple and quadruple infections may be a rare occurrence in animals, although common in tsetse flies (Njiru et al 2004). The use of universal tests for pathogenic trypanosomes would reduce the cost of PCR three to five times as the number of reactions required per sample would be reduced to one (Desquesnes and Davila 2002).…”

Section: Discussionmentioning

confidence: 96%

The use of ITS1 rDNA PCR in detecting pathogenic African trypanosomes

Njiru

Constantine

Guya³

et al. 2004

Parasitol Res

239

225

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There are 11 different pathogenic trypanosomes in trypanosomiasis endemic regions of Africa. Their detection and characterisation by molecular methods relies on species-specific primers; consequently several PCR tests have to be made on each sample. Primers ITS1 CF and ITS1 BR, previously designed to amplify the internal transcribed spacer (ITS1) of rDNA, have been evaluated for use in a universal diagnostic test for all pathogenic trypanosomes. Blood was collected from 373 cattle and 185 camels. The primers gave constant PCR products with the stocks of each taxon tested. Members of subgenus Trypanozoon (T. brucei brucei, T. evansi, T. b. rhodesiense and T. b. gambiense) gave a constant product of approximately 480 bp; T. congolense, savannah 700 bp, T. congolense kilifi 620 bp and T. congolense forest 710 bp: T. simiae 400 bp, T. simiae tsavo 370 bp, T. godfreyi 300 bp and T. vivax 250 bp. The sensitivity of the test ranged from 10 pg for Trypanozoon, T. congolense clade and T. vivax to 100 pg for T. simiae and T. godfreyi. The primers detected cases of multi-taxa samples, although the sensitivity was reduced with an increase in the combinations. A better detection rate of trypanosome DNA was recorded with buffy coats than from direct blood. With the field samples, the diagnostic sensitivity was close to the sensitivity obtained using single reactions with species-specific primers for Trypanozoon 38/40 (95%) and T. congolense savannah 30/33 (90.9%) but was lower with T. vivax 25/31 (77.4%). The primers offer promise as a routine diagnostic tool through the use of a single PCR; however, further evaluation is recommended.

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Section: Discussionmentioning

confidence: 96%

The use of ITS1 rDNA PCR in detecting pathogenic African trypanosomes

Njiru

Constantine

Guya³

et al. 2004

Parasitol Res

239

225

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“…With the development of molecular biology during the last decades, specific DNA sequences of different trypanosome species and subspecies have been identified and several PCR based methods were developed to improve the detection of various parasites. Applied in human and animal trypanosomosis, PCR appeared as a reliable, sensitive and specific techniques enabling to detect different trypanosome species and subspecies in vertebrate hosts as well as in tsetse flies (Masiga et al , 1992; Desquesnes & Tresse, 1996; Penchenier et al , 2000; Mugutu et al , 2001; Picozzi et al , 2002; Geysen et al , 2003; Delespaux et al , 2003; Gonzales et al , 2005; Cox et al , 2005). …”

Section: Introductionmentioning

confidence: 99%

Trypanosoma vivax,T. congolense“forest type” andT. simiae: prevalence in domestic animals of sleeping sickness foci of Cameroon

et al. 2011

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In order to better understand the epidemiology of Human and Animal trypanosomiasis that occur together in sleeping sickness foci, a study of prevalences of animal parasites (Trypanosoma vivax, T. congolense “forest type”, and T. simiae) infections was conducted on domestic animals to complete the previous work carried on T. brucei gambiense prevalence using the same animal sample. 875 domestic animals, including 307 pigs, 264 goats, 267 sheep and 37 dogs were sampled in the sleeping sickness foci of Bipindi, Campo, Doumé and Fontem in Cameroon. The polymerase chain reaction (PCR) based method was used to identify these trypanosome species. A total of 237 (27.08%) domestic animals were infected by at least one trypanosome species. The prevalence of T. vivax, T. congolense “forest type” and T. simiae were 20.91%, 11.42% and 0.34% respectively. The prevalences of T. vivax and T. congolense “forest type” differed significantly between the animal species and between the foci (p < 0.0001); however, these two trypanosomes were found in all animal species as well as in all the foci subjected to the study. The high prevalences of T. vivax and T. congolense “forest type” in Bipindi and Fontem-Center indicate their intense transmission in these foci.

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“…Furthermore, DNA-based methods for diagnosis have generally facilitated epidemiological studies of trypanosomes. 22 However, in the Philippines, the use of PCR technology is currently limited to academic and specialized government institutions. The possibility of field applications is currently being hampered by economic issues and questions, especially pertaining to its suitability as a diagnostic test in the field or countryside.…”

Section: Resultsmentioning

confidence: 99%

Parasitological examination for Trypanosoma theileri infection of cattle from Quirino Province, Philippines

Fernandez¹,

Baticados²,

Baticados³

2010

VMRR

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Abstract:Trypanosoma theileri is one of the protozoan parasites reported in cattle and carabaos in the Philippines. The distribution of T. theileri infection in livestock in this country is not well established. To date, more than two decades have passed without any new T. theileri prevalence reported in the country. The present study endeavored to determine whether the parasite is endogenously present in cattle from the remote areas of Region II (Cagayan Valley), particularly in the province of Quirino, Philippines and to establish the current parasite prevalence of T. theileri in this province using the blood parasite examination method. A total of 246 field blood samples of cattle was collected from 5/6 (83%) of the municipalities of Quirino Province, Philippines. Blood parasite examination of Giemsa-stained smears revealed that all samples were negative for the presence of the T. theileri parasite. Given that the mainstay of parasitological examination in the field is the classic blood parasite examination method, the data suggest that the negative results of these examinations can probably be attributed to very low parasitemia.

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Application of PCR and DNA probes in the characterisation of trypanosomes in the blood of cattle in farms in Morogoro, Tanzania

Cited by 25 publications

References 28 publications

The use of ITS1 rDNA PCR in detecting pathogenic African trypanosomes

The use of ITS1 rDNA PCR in detecting pathogenic African trypanosomes

Trypanosoma vivax,T. congolense“forest type” andT. simiae: prevalence in domestic animals of sleeping sickness foci of Cameroon

Parasitological examination for Trypanosoma theileri infection of cattle from Quirino Province, Philippines

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