Eimeria maxima: the influence of host genotype on parasite reproduction as revealed by quantitative real-time PCR.
The influence of host genotype on susceptibility to infection with Eimeria species has long been recognised, but beyond monitoring pathological severity or magnitude of oocyst excretion attempts to quantify fluctuations in parasite reproduction within the host have previously relied upon labour-intensive microscopic analysis. The development and application of a quantitative real-time PCR assay has opened this biological 'black box', permitting the sensitive and reproducible enumeration of parasite genomes throughout the course of infection. Generic and species-specific quantitative PCR methods are described, based upon the conserved 5S ribosomal RNA coding sequence of nine avian and murine Eimeria species and the Eimeria maxima MIC1 gene, respectively. These complementary assays have been applied to study the influence of host genotype on resistance to infection with E. maxima, revealing significant differences in parasite load between 'resistant' Line C and 'susceptible' Line 15I inbred chickens 5 days after infection. Parasite DNA remained detectable up to 20 days post-infection; 11 days after the last oocysts had been detected leaving the host.
Blake
Damer P
DP
Enteric Immunology Group, Institute for Animal Health, Compton, Nr. Newbury, Berkshire RG20 7NN, UK.
Hesketh
Patricia
P
Archer
Andrew
A
Shirley
Martin W
MW
Smith
Adrian L
AL
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D016207:Cytokines / Q000235:genetics; D004261:DNA Replication / Q000235:genetics; D016054:DNA, Protozoan / Q000032:analysis; D004198:Disease Susceptibility; D004539:Eimeria / Q000235:genetics*; D005838:Genotype; D055436:Growth Differentiation Factor 15; D006790:Host-Parasite Interactions / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D033661:Oocysts / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D016415:Sequence Alignment / Q000379:methods; D013045:Species Specificity
D016207:Cytokines; D016054:DNA, Protozoan; C526141:Gdf15 protein, mouse; D055436:Growth Differentiation Factor 15; D012341:RNA, Ribosomal, 5S