PCR identification of chicken Eimeria: A simplified read-out

Schnitzler, Beate E.; Thebo, P.; Tomley, Fiona M.; Uggla, A.; Shirley, M. W.

doi:10.1080/03079459995091

Cited by 88 publications

(63 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…33). ITS sequences often are used for molecular diagnostics and phylogenetic analyses because of the sensitivity and specificity provided by their high copy number and relatively rapid evolution, although intraclonal polymorphism has proven limiting (33,34). Here, multiplex SNP genotyping for E. tenella provided far greater genetic power.…”

Section: Discussionmentioning

confidence: 99%

Population, genetic, and antigenic diversity of the apicomplexanEimeria tenellaand their relevance to vaccine development

Blake

Clark

Macdonald

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

105

102

View full text Add to dashboard Cite

The phylum Apicomplexa includes serious pathogens of humans and animals. Understanding the distribution and population structure of these protozoan parasites is of fundamental importance to explain disease epidemiology and develop sustainable controls. Predicting the likely efficacy and longevity of subunit vaccines in field populations relies on knowledge of relevant preexisting antigenic diversity, population structure, the likelihood of coinfection by genetically distinct strains, and the efficiency of cross-fertilization. All four of these factors have been investigated for Plasmodium species parasites, revealing both clonal and panmictic population structures with exceptional polymorphism associated with immunoprotective antigens such as apical membrane antigen 1 (AMA1). For the coccidian Toxoplasma gondii only genomic diversity and population structure have been defined in depth so far; for the closely related Eimeria species, all four variables are currently unknown. Using Eimeria tenella, a major cause of the enteric disease coccidiosis, which exerts a profound effect on chicken productivity and welfare, we determined population structure, genotype distribution, and likelihood of crossfertilization during coinfection and also investigated the extent of naturally occurring antigenic diversity for the E. tenella AMA1 homolog. Using genome-wide Sequenom SNP-based haplotyping, targeted sequencing, and single-cell genotyping, we show that in this coccidian the functionality of EtAMA1 appears to outweigh immune evasion. This result is in direct contrast to the situation in Plasmodium and most likely is underpinned by the biology of the direct and acute coccidian life cycle in the definitive host.Eimeria | coccidiosis | population structure | chickens | food security

show abstract

Section: Discussionmentioning

confidence: 99%

Population, genetic, and antigenic diversity of the apicomplexanEimeria tenellaand their relevance to vaccine development

Blake

Clark

Macdonald

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

105

102

View full text Add to dashboard Cite

show abstract

“…This test was found to be highly sensitive and could be used to differentiate between the seven species of Eimeria. [40][41][42] However, the method required a different primer pair for each species, resulting in seven separate PCR reactions to test a given sample, and was therefore deemed labor-intensive and time-consuming. Additionally, because this approach gave limited information about strain variation, further work was carried out to develop a test based on the use of the second ITS region.…”

Section: Targeted Pcr On Individual Speciesmentioning

confidence: 99%

Designing strategies for the control of coccidiosis in chickens on poultry farms using modern diagnostic tools

Frölich¹,

Farhat²,

Wallach³

2013

RIP

View full text Add to dashboard Cite

Abstract:Coccidiosis is caused by the intracellular protozoan parasite Eimeria and is a major worldwide problem in the poultry industry today. The current strategies to control the disease still rely heavily on anticoccidial drugs and on the use of shuttle programs to periodically introduce attenuated live parasites onto poultry farms. Recently, because of the improved performance of broiler chickens, farmers are able to raise chickens for market within a period of only 5-6 weeks. This short growth period enables the development of strategies for coccidiosis control either by ensuring an even exposure of chickens to parasites in the litter, inducing immunity against reinfection, or by keeping parasite numbers below a threshold level wherein only subclinical infections will occur. Therefore, the development and use of new diagnostic tools to precisely identify the Eimeria species and strain and to measure the number of oocysts in the feces and litter can be very helpful in predicting exposure and designing strategies for the control of coccidiosis in the field. The purpose of this study was to describe recently developed methods of diagnosis in terms of their sensitivity and usefulness, describe ways in which litter oocyst numbers can be quantitated, and start to develop an analytical tool that can be used for designing strategies for the control of coccidiosis. A multiplex polymerase chain reaction-based assay has been developed that enables the identification of all seven Eimeria species in a single reaction tube. Previous reports have shown it is possible to detect one oocyst using a polymerase chain reaction-based assay; however, other reports have found that under both laboratory and field conditions the minimal level of sensitivity is only 20 oocysts. In addition, the present authors' studies have found that when using spiked mixed samples, the level of detection of a contaminating Eimeria species is only 5%. Therefore, it is imperative that a new method be developed with a tenfold higher level of sensitivity for field-testing. Once this test is developed and validated, it can be used to monitor the litter and assess the parameters (climate, hygiene, breed of chicken, etc) that govern coccidial outbreaks on farms.

show abstract

“…Several reports described the development of PCR assays for species discrimination of Eimeria spp. using as targets different regions of the ribosomal cistrons, including the 5S rRNA (Stucki et al 1993), the small subunit rRNA (Tsuji et al 1997), the ribosomal internal transcribed spacer 1 (ITS-1) (Schnitzler et al 1998(Schnitzler et al , 1999, and ITS-2 (Woods et al 2000;Gasser et al 2001). …”

Section: Introductionmentioning

confidence: 99%

Multiplex PCR assay using SCAR primers to detect Eimeria spp. in chicken

et al. 2012

View full text Add to dashboard Cite

About 11 faecal samples from various regions of Tamil Nadu and Andhra Pradesh containing mixed spectrum of Eimeria species detected by morphometry viz. 15.4 9 11.2, 28.5 9 20.3, 31.1 9 18.5, 13.2 9 12.4, 20.8 9 17.5 and 22 9 18 lm for E. acervulina, E. brunetti, E. maxima, E. mitis, E. necatrix, E. praecox and E. tenella respectively were taken for the study. The oocysts were concentrated and purified using faecal harvest protocol. The genomic DNA is extracted as per kit protocol. The amplicons of sizes 539 bp (E. tenella) and 460 bp (E. mitis) obtained could be visualised in a single lane for the multiplex PCR assay using sequence characterized amplified region primers.

show abstract

PCR identification of chicken Eimeria: A simplified read-out

Cited by 88 publications

References 7 publications

Population, genetic, and antigenic diversity of the apicomplexanEimeria tenellaand their relevance to vaccine development

Population, genetic, and antigenic diversity of the apicomplexanEimeria tenellaand their relevance to vaccine development

Designing strategies for the control of coccidiosis in chickens on poultry farms using modern diagnostic tools

Multiplex PCR assay using SCAR primers to detect Eimeria spp. in chicken

Contact Info

Product

Resources

About