Sensitivity of different Trypanosoma vivax specific primers for the diagnosis of livestock trypanosomosis using different DNA extraction methods

Gonzáles, José L.; Loza, A.; Chacón, Eduardo

doi:10.1016/j.vetpar.2005.10.024

Cited by 21 publications

(11 citation statements)

References 13 publications

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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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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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“…The development of practical techniques to prepare DNA samples are widely studied for applied in epidemiological studies such as for forensic medicine (Jones et al 1999, Gonzales et al 2006, Lin et al 2006. In this study, PCR reactions were prepared with DNA samples extracted directly from blood clots.…”

Section: Introductionmentioning

confidence: 99%

Anaplasma marginale infection in cattle from south-western Amazonia

Brito¹,

Oliveira²,

Rocha³

et al. 2010

Pesq. Vet. Bras.

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249Pesq. Vet. Bras. 30(3):249-254, março 2010 RESUMO.-[Infecção por Anaplasma marginale em bovinos na Amazônia Sul Ocidental, Brasil.] O presente estudo fornece os primeiros dados epidemiológicos relativos a infecção por Anaplasma marginale em bovinos criados na Amazônia Sul Ocidental brasileira. Foi adaptado um procedimento simples para a extração de DNA a partir de coágulos sanguíneos coletados em sete microrregiões do estado de Rondônia e duas mesoregiões do estado do Acre. A técnica da reação em cadeia da polimerase (PCR) foi aplicada para avaliar a freqüência da infecção por A. marginale em bovinos com idade entre 4 e 12 meses. Após a extração do DNA de cada amostra, a infecção nos bovinos foi investigada pela amplificação do gene "msp5" de A. marginale. As técnicas de amplificação do DNA revelaram que a freqüência de infecção por A. marginale foi de 98,6% (1.627/1.650) nas amostras provenientes de Rondônia e de 92,87% (208/225) nas amostras do Acre. A alta freqüên-cia da infecção por A. marginale nos animais com idade entre 4 e 12 meses indica uma situação de estabilidade enzoótica nas regiões estudadas, as quais são compará-veis às detectadas por técnicas de imunodiagnóstico em outras regiões endêmicas no Brasil. A extração do DNA através do método aqui descrito pode ser utilizado em estudos epidemiológicos sobre a anaplasmose bovina e outros hemoparasitas. The present study provides the first epidemiological data regarding infection by Anaplasma marginale in cattle reared in south-western Brazilian Amazonia. One simple procedure was adapted for the extraction of DNA from blood clots collected in seven microregions of Rondônia State and two mesoregions of Acre State. PCR method was used to asses the frequency of A. marginale infections in 4 to12-month-old cattle. The cattle infection was investigated by polymerase chain reaction (PCR) using the specific primer "msp5" for A. marginale. The DNA amplifications revealed that the mean frequency of A. marginale infection was 98.6% (1,627/1,650) in samples from Rondonia, and 92.87% (208/225) in samples from Acre. The high frequency of A. marginale infections in 4 to 12-month-old cattle indicate a situation of enzootic stability in the studied areas and are comparable to those detected by immunodiagnosis in different endemic regions in Brazil. The DNA extraction of clotted blood method described here can be used for epidemiological studies on anaplasmosis and other bovine hemoparasites.

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“…For a 3-mm Whatman filter paper method, the sensitivity was 100%, and for methanol and Chelex methods, it was 73% and 93%, respectively. Gonzalez et al [23] reported the use of different DNA extraction techniques, and they found that WBC lysis with proteinase K and heat inactivation was the best technique. Whereas, FTA cards performed better than untreated filter papers, and were nearly comparable to the sensitivity of WBC lysis.…”

Section: Discussionmentioning

confidence: 99%

Imported Malaria in United Arab Emirates: Evaluation of a New DNA Extraction Technique Using Nested PCR

et al. 2009

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Local malaria transmission in the United Arab Emirates (UAE) came to an end in 1997. Nevertheless, UAE has been subjected to substantial importation of malaria cases from abroad, concerning both UAE nationals and immigrants from malarious countries with a total number of 2,119 cases in 2007. To evaluate a new DNA extraction technique using nested PCR, blood samples were collected from 132 individuals who presented to Infectious Diseases Department in Rashid Hospital, Dubai, and Central Department of Malaria Control with fever and persistent headache. Giemsa-stained blood films and ELISA test for malaria antibodies were carried out for detection of Plasmodium infection. Plasmodium infections were identified with the genus-specific primer set and species differentiation using nested PCR. A rapid procedure for diagnosis of malaria infections directly from dried blood spots using for the first time DNA extract from FTA Elute cards was evaluated in contrast to extraction techniques using FTA classic cards and rapid boiling technique. Our new simple technique for DNA extraction using FTA Elute cards was very sensitive giving a sensitivity of 100% compared to 94% using FTA classic cards and 62% in the rapid boiling technique. No complex preparation of blood samples was required prior to the amplification. The production cost of DNA isolation in our PCR assay was much less in comparable to that of other DNA extraction protocols. The nested PCR detected plasmodial infection and could differentiate P. falciparum from P. vivax, and also detected the mixed infection.

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Sensitivity of different Trypanosoma vivax specific primers for the diagnosis of livestock trypanosomosis using different DNA extraction methods

Cited by 21 publications

References 13 publications

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

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

Anaplasma marginale infection in cattle from south-western Amazonia

Imported Malaria in United Arab Emirates: Evaluation of a New DNA Extraction Technique Using Nested PCR

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