Direct and Sensitive Detection of Trypanosoma Evansi by Polymerase Chain Reaction

Omanwar, Swati; Rao, J. R.; Butchaiah, G.; Basagoudanavar, Suresh H.; Singh, Rajkumar

doi:10.1556/avet.47.1999.3.9

Cited by 23 publications

(6 citation statements)

References 10 publications

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“…Trypanozoon specific primers have been designed previously: TBR primers which target a 177 bp repeat [4], pMUTEC primers targeting a retrotransposon [5] and ORPHON primers which target the spliced leader sequence [6]. Most of them have been tested on cattle [7,8], water buffaloes [9] or goats [10]. PCR tests for diagnosis of T. congolense and T. vivax infections exist as well [11].…”

Section: Introductionmentioning

confidence: 99%

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Claes

Radwanska

Urakawa

et al. 2004

Kinetoplastid Biol Dis

103

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BackgroundBased on the recently sequenced gene coding for the Trypanosoma evansi (T. evansi) RoTat 1.2 Variable Surface Glycoprotein (VSG), a primer pair was designed targeting the DNA region lacking homology to other known VSG genes. A total of 39 different trypanosome stocks were tested using the RoTat 1.2 based Polymerase Chain Reaction (PCR).ResultsThis PCR yielded a 205 bp product in all T. evansi and in seven out of nine T. equiperdum strains tested. This product was not detected in the DNA from T. b. brucei, T. b. gambiense, T. b. rhodesiense, T. congolense, T. vivax and T. theileri parasites. The Rotat 1.2 PCR detects as few as 10 trypanosomes per reaction with purified DNA from blood samples, i.e. 50 trypanosomes/ml.ConclusionPCR amplification of the RoTat 1.2 VSG gene is a specific marker for T. evansi strains, except T. evansi type B, and is especially useful in dyskinetoplastic strains where kDNA based markers may fail to amplify. Furthermore, our data support previous suggestions that some T. evansi stocks have been previously misclassified as T. equiperdum.

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Section: Introductionmentioning

confidence: 99%

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Claes

Radwanska

Urakawa

et al. 2004

Kinetoplastid Biol Dis

103

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“…The specificity studies indicated that the primary specific 821-bp and the nested 270-bp PCR products were not amplified even from concentrations of 10 ng of DNA from other blood parasite including Thieleria annulata, Babesia bigemina and blood cells from non infected mice and camels under the same stringency condition described in this study. Based on these sensitivity and specificity results of this study, the described nPCR should be considered as the most highly sensitive and specific assay compared to the previously described PCR-based detection assays [ 10 , 12 , 13 , 19 ]. The second amplification step using the nested primers TE3 and TE4 is necessary to confirm the specificity of the primary amplified product and to increase the sensitivity of the PCR-based assay by at least 1000 times particularly, when the concentration of the T. evansi DNA in the sample is less than 10 pg.…”

Section: Discussionmentioning

confidence: 86%

“…The economic importance of T. evansi infection is mainly attributed to clinical disease in camels in many parts of the world [ 14 - 18 ]. PCR has been used successfully in detecting infection with T. evansi in buffaloes [ 12 , 19 ], horses [ 13 ] and in camels [ 11 ]. So far, there is no comprehensive data on the use of PCR for detecting infection in Sudanese breed of dromedary camels ( Camelus dromedarius ).…”

Section: Discussionmentioning

confidence: 99%

A simple and rapid method for detection of Trypanosoma evansiin the dromedary camel using a nested polymerase chain reaction

Aradaib

Majid

2006

Kinetoplastid Biol Dis

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A nested polymerase chain reaction (nPCR)-based assay, was developed and evaluated for rapid detection of Trypanosoma evansi in experimentally infected mice and naturally infected camels (Camelus dromedarius). Four oligonucleotide primers (TE1, TE2, TE3 and TE4), selected from nuclear repetitive gene of T. evansi, were designed and used for PCR amplifications. The first amplification, using a pair of outer primers TE1 and TE2, produced a 821-bp primary PCR product from T. evansi DNA. The second amplification, using nested (internal) pair of primers TE3 and TE4, produced a 270-bp PCR product. T. evansi DNAs extracted from blood samples of experimentally infected mice and naturally infected Sudanese breed of dromedary camels were detected by this nested PCR-based assay. The nested primers TE3 and TE4 increased the sensitivity of the PCR assay and as little as 10 fg of T. evansi DNA (equivalent to a single copy of the putative gene of the parasite) was amplified and visualized onto ethidium bromide-stained agarose gels. Amplification products were not detected when the PCR-based assay was applied to DNA from other blood parasites including Thieleria annulata, Babesia bigemina or nucleic acid free samples. Application of this nPCR-based assay to clinical samples resulted in direct detection of T. evansi from a variety of tissue samples collected from experimentally infected mice and blood from naturally infected camels. The described nPCR-based assay provides a valuable tool to study the epidemiology of T. evansi infection in camels and other susceptible animal populations.

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“…The further confirmation of the identity of the candidate trypanosome by PCR through using primers that specifically targeted the ITS1 region of the rDNA gene of T. evansi that is performed in the present study, is similar to the result reported by Croof [50] who used molecular method (PCR) in his study of 40 camels which were tested parasitologically and serologically where 90% of them were found to be positive. PCR has been used in detection of infection with T. evansi in buffaloes [51][52] , in horses [53] and in camels [54] . There was no comprehensive data on the use of PCR for detection of infection in Sudanese breed of dromedary camels (Camelus dromedarius).…”

Section: Discussionmentioning

confidence: 99%

Investigation of Drug Susceptibility in Rats Experimentally Infected with Trypanosoma evansi Isolated from Camels in Sudan

Abuessailla¹,

Aa²,

Agab³

et al. 2017

Int. J. Life. Sci. Scienti. Res.

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A number of 18 adults male outbred albino rats, weighing between 133-137g were used to investigate the drug susceptibility of Trypanosoma evansi strain isolated from naturally infected dromedary camels in Umbadir area, North Kordofan State, Sudan. The rats were divided into 3 groups (C, D and F) of 6 animals each. Group C and D were infected intraperitoneally with T. evansi (Umbadir stabilate) with 1×10 4 Trypanosome for the inoculum. Group D rats were given quinapyramine sulphate (20 mg/Kg bwt) after parasitaemia was evident. Group F was left as healthy uninfected control for the stabilate. When parasite counts were one or more parasites per field, counting in haemocytometer were used for exact number of parasite per cubic millimeter using Neubaeur's counter. Parasites from tail blood were first fixed, stained and diluted in trypanosome diluting reagent. The parasites were diluted to the level that can be easily counted in WBC counting chamber in the haemocytometer. The total number of parasites was expressed as log 10 number of parasites per ml of blood. The presence and degree of parasitaemia were determined daily for each rat by examining tail blood. The identity of the local stabilates of Trypanosoma evansi was confirmed through adopting PCR where primers that target the internal transcribed spacer one (ITS1) of the ribosomal DNA were used. There was significant reduction in serum glucose and potassium as well as significant increase in total protein, urea, calcium, albumin and cholesterol in group C. The Umbadir stabilate showed low mortality and high sensitivity to quinapyramine sulphate.

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Direct and Sensitive Detection of Trypanosoma Evansi by Polymerase Chain Reaction

Cited by 23 publications

References 10 publications

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A simple and rapid method for detection of Trypanosoma evansiin the dromedary camel using a nested polymerase chain reaction

Investigation of Drug Susceptibility in Rats Experimentally Infected with Trypanosoma evansi Isolated from Camels in Sudan

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