Pentaplex PCR assay for rapid differential detection of Babesia bigemina, Theileria annulata, Anaplasma marginale and Trypanosoma evansi in cattle

Biologicals ◽  
2020 ◽  
Vol 63 ◽  
pp. 81-88 ◽  
Author(s):  
Anita Ganguly ◽  
Biswa Ranjan Maharana ◽  
Indrajit Ganguly
Keyword(s):  
Anaplasma Marginale ◽  
Trypanosoma Evansi ◽  
Theileria Annulata ◽  
Differential Detection ◽  
Babesia Bigemina ◽  
Pcr Assay

Molecular Detection, Phylogenetic Analysis, and Genetic Diversity of Theileria annulata, Babesia bigemina, and Anaplasma marginale in Cattle in Three Districts of Egypt

2020 ◽  
Vol 65 (3) ◽  
pp. 620-627
Author(s):  
Khaled Mohamed El-Dakhly ◽  
Waleed M. Arafa ◽  
Saad Soliman ◽  
Omima Ramadan Abdel-Fatah ◽  
Ahmed Anwar Wahba ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Phylogenetic Analysis ◽  
Molecular Detection ◽  
Anaplasma Marginale ◽  
Theileria Annulata ◽  
Babesia Bigemina

scholarly journals Epidemiology, haematology and molecular characterization of haemoprotozoon and rickettsial organisms causing infections in cattle of Jammu region, North India

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Rabjot Kaur ◽  
Anish Yadav ◽  
Shafiya I. Rafiqi ◽  
Rajesh Godara ◽  
Vikrant Sudan ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Genetic Characterization ◽  
Disease Mapping ◽  
Anaplasma Marginale ◽  
Theileria Annulata ◽  
Genomic Diversity ◽  
North India ◽  
Babesia Bigemina ◽  
Prevalence Studies

Abstract Background The present study was aimed at establishing the prevalence, epidemiology and molecular characterization of major haemoprotozoons (Babesia and Theileria) and rickettsia (Anaplasma) of cattle in Jammu region (North India) using microscopy and Polymerase Chain Reaction (PCR). Hematology, microscopy and PCR based prevalence studies were undertaken with 278 whole blood samples from cattle. Molecular prevalence studies were followed by genetic characterization of the isolates of Babesia, Anaplasma and Theileria spp. based on 18S rRNA, 16S rRNA and Tams1 gene, respectively. The data related to metrology and epidemiological variables like temperature, rainfall, season, age and type of livestock rearing was analyzed and correlated with occurrence of disease by statistical methods. Results The prevalence based on microscopy was 12.9% (36/278) whereas PCR recorded 30.22% (84/278) animals positive for haemoparasitic infections. All the samples found positive by microscopy were also recorded positive by PCR. Thus the study revealed prevalence of Babesia bigemina, Anaplasma marginale and Theileria annulata to be 9.7, 16.5 and 0.7% respectively. The metrological and epidemiological variables made inroads for the propagation of vector ticks and occurrence of infection. Haematological alterations predominantly related to decrease in haemoglobin, red blood cell count and packed cell volume were evident in diseased animals and collaterally affected the productivity. Further the genetic characterization of Babesia bigemina. (MN566925.1, MN567603, MN566924.1), Anaplasma marginale. (MH733242.1, MN567602.1) and Theileria annulata (MT113479) provided a representative data of the isolates circulating in the region and their proximity with available sequences across the world. Conclusions Despite holding much significance to the animal sector, comprehensive disease mapping has yet not been undertaken in several parts of India. The present study provides a blue print of disease mapping, epidemiological correlations and genomic diversity of Babesia bigemina, Anaplasma marginale and Theileria annulata circulating in the region.

A comparative analysis of microscopy and PCR based detection methods for Babesia and Trypanosoma infecting bovines and assessment of risk factors

2018 ◽  
Author(s):  
B. R. Maharana ◽  
B. Kumar ◽  
J. P. Joseph ◽  
T. K. Patbandha
Keyword(s):  
Risk Factors ◽  
Odds Ratio ◽  
Regression Models ◽  
Trypanosoma Evansi ◽  
Detection Methods ◽  
Babesia Bigemina ◽  
Multivariate Logistic Regression ◽  
Pcr Assay ◽  
Logistic Regression Models ◽  
The Status

The present study was carried out to evaluate and compare the status of infection and assessment of risk factors in 353 blood samples (144 cattle and 209 buffaloes) of bovines by PCR assay along with microscopic examinations (ME). ME revealed prevalence of Babesia bigemina and Trypanosoma evansi to be 22.91% and 0.69%, respectively in cattle and 12.44% and 0.95%, respectively in buffaloes. Conversely, PCR assay was able to detect 40.97% and 3.47 % prevalence of B. bigemina and T. evansi in cattle and 23.92% and 6.69% in buffaloes, respectively. The result revealed that the PCR assay was 100% sensitive and 82.9% specific when compared with ME for babesiosis and 100% sensitive and 95.42% specific for trypanosomosis. Multivariate logistic regression models showed that risk of babesiosis was significantly higher in cattle (Odds ratio (OR) =2.207, P=0.001) compared to buffaloes. The risk for surra in male buffaloes increased by 6.37 times (OR= 6.375, P=0.013). Conversely, risk of babesiosis was significantly lower in male cattle than females (OR= 0.467, P=0.044).

scholarly journals Séroprévalence et facteurs de risque des hémoparasitoses (theilériose, babésiose et anaplasmose) chez les bovins dans quatre grandes régions d’élevage du Maroc

2015 ◽  
Vol 67 (4) ◽  
pp. 235 ◽  
Author(s):  
T. Rahali ◽  
H. Sahibi ◽  
A. Sadak ◽  
S. Ait Hamou ◽  
B. Losson ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Facteurs De Risque ◽  
Anaplasma Marginale ◽  
Theileria Annulata ◽  
Enzyme Linked Immunosorbent Assay ◽  
Babesia Bigemina ◽  
Infections Mixtes

Les hémoparasitoses (theilériose, babésiose et anaplasmose) sont des maladies vectorielles graves dont l’impact économique et sur la santé des élevages bovins au Maroc est considérable. L’objectif de cette étude, réalisée dans quatre régions d’élevage bovin laitier, a été de déterminer la séroprévalence et les facteurs de risque de ces infections. Au total 1 040 bovins ont été examinés dans 96 exploitations. La technique competitive enzyme-linked immunosorbent assay (cELISA) et la réaction d’immunofluorescence indirecte ont été utilisées pour la détermination de la séroprévalence, et le test statistique V de Cramer a permis d’analyser les facteurs de risque d’infection des bovins par les hémoparasites. Sur 1 040 sérums analysés, 689 (66,3 p. 100) ont été séropositifs ; la séroprévalence a varié selon le parasite considéré : Theileria annulata (35,4 p. 100), Anaplasma marginale (20,5 p. 100), Babesia bigemina (13,6 p. 100) et B. bovis (12,0 p. 100). Par ailleurs, 155 sérums (14,9 p. 100) ont présenté des infections mixtes (deux ou trois hémoparasites simultané- ment). L’analyse statistique a montré que l’âge (C = 0,62) et le type d’élevage (C = 0,60) ont eu une forte influence sur la séroprévalence, tandis que la race (C = 0,46) et le sexe (C = 0,29) ont eu une influence moindre. Le climat (C = 0,06) n’a pas semblé avoir d’impact. Ces résultats aideront à comprendre la situation épizootique des hémoparasitoses vectorielles chez les bovins afin d’établir une stratégie adéquate pour leur contrôle.

scholarly journals Development of a multiplex PCR assay for simultaneous detection of Theileria annulata, Babesia bovis and Anaplasma marginale in cattle

2013 ◽  
Vol 133 (2) ◽  
pp. 222-229 ◽  
Author(s):  
Huseyin B. Bilgiç ◽  
Tülin Karagenç ◽  
Martin Simuunza ◽  
Brian Shiels ◽  
Andy Tait ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Anaplasma Marginale ◽  
Theileria Annulata ◽  
Babesia Bovis ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals Utilisation d’un test multiple basé sur la réaction de polymérase en chaîne pour des enquêtes épidémiologiques sur les hémoparasites des bovins au Mexique

1993 ◽  
Vol 46 (1-2) ◽  
pp. 71-75
Author(s):  
Julio Vicente Figueroa ◽  
J.A. Alvarez ◽  
J.A. Ramos ◽  
C.A. Vega ◽  
G.M. Buening
Keyword(s):  
Anaplasma Marginale ◽  
Babesia Bigemina ◽  
Dot Blot ◽  
Multiple Pcr

Une étude a été effectuée sur la possibilité d'appliquer la technique de la réaction de polymérase en chaîne (PCR) pour la détection simultanée des hémoparasites bovins Babesia bigemina, B. bovis et Anaplasma marginale. Des échantillons de sang de bovins ont été récoltés dans des ranches d'une zone endémique identifiée au préalable dans la péninsule de Yucatan au Mexique, et ont été préparés pour analyse par PCR. Ils ont été soumis à l'amplification de l'ADN dans un tube à réaction contenant des amorces d'oligonucléotides spécifiques pour l'ADN de chaque espèce d'hémoparasite. Les produits de la PCR ont été détectés par hybridation en Dot-Blot de l'acide nucléique utilisant des sondes d'ADN non-radioactives, spécifiques, marquées à la digoxigénine par PCR. 420 échantillons analysés par le test multiple PCR-sonde ADN ont montré des taux de prévalence de 66,7 %, 60,1 et 59,6 % pour B. bigemina, B. bovis et A. marginale, respectivement. L'analyse multiple par PCR a montré que des animaux ayant des infections simples, doubles ou triples pouvaient être détectés par les sondes d'ADN spécifiques. La procédure est proposée comme un outil de valeur pour l'analyse épidémiologique dans les régions où ces espèces d'hémoparasites infectent les bovins simultanément.

scholarly journals Multiplex real-time reverse transcription polymerase chain reaction for differential detection of H5, N1, and N8 genes of highly pathogenic avian influenza viruses

2017 ◽  
Vol 62 (No. 4) ◽  
pp. 211-220 ◽  
Author(s):  
YR Park ◽  
EM Kim ◽  
YJ Lee ◽  
SG Yeo ◽  
CK Park
Keyword(s):  
Polymerase Chain Reaction ◽  
Avian Influenza ◽  
Highly Pathogenic Avian Influenza ◽  
Differential Detection ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Highly Pathogenic ◽  
Pathogenic Avian Influenza ◽  
Polymerase Chain ◽  
H5 Subtype

Rapid and differential diagnosis of highly pathogenic avian influenza virus (HPAIV) subtype H5 is essential for the effective prevention and control of outbreaks caused by this pathogen. In this study, we describe a one-step multiplex real-time reverse transcription polymerase chain reaction (mRRT-PCR), using H5-, N1-, and N8-specific primers and probes, for differential detection of two HPAIVs (H5N1 and H5N8) and other H5-subtype AIVs. Using the mRRT-PCR assay, we were able to detect H5N1, H5N8, and other H5-subtype AIVs in a one-tube reaction, with high specificity; furthermore, using an in silico PCR program, we confirmed that this assay can detect nearly all H5, N1, and N8 genes of AIVs currently available in the Influenza Sequence Database. The limit of detection of the assay was determined to be as low as 100 copies/reaction for each target gene, and was comparable to limits of detection of previously reported mRRT-PCR assays. Thus, the mRRT-PCR assay described here can serve as a rapid and reliable differential diagnostic tool for the monitoring and surveillance of H5N1, H5N8, and other H5-subtype AIVs in countries where these pathogens are problematic.

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