Adequately sensitive and specific methods to diagnose pertussis in adolescents and adults are not widely available. Currently, no Food and Drug Administration-approved diagnostic assays are available for the serodiagnosis of Bordetella pertussis. Since concentrations of B. pertussis-specific antibodies tend to be high during the later phases of disease, a simple, rapid, easily transferable serodiagnostic test was developed. This article describes test development, initial evaluation of a prototype kit enzyme-linked immunosorbent assay (ELISA) in an interlaboratory collaborative study, and analytical validation. The data presented here demonstrate that the kit met all prespecified criteria for precision, linearity, and accuracy for samples with anti-pertussis toxin (PT) immunoglobulin G (IgG) antibody concentrations in the range of 50 to 150 ELISA units (EU)/ml, the range believed to be most relevant for serodiagnosis. The assay met the precision and linearity criteria for a wider range, namely, from 50 to 200 EU/ml; however, the accuracy criterion was not met at 200 EU/ml. When the newly adopted World Health Organization International Standard for pertussis antiserum (human) reference reagent was used to evaluate accuracy, the accuracy criteria were met from 50 to 200 international units/ml. In conclusion, the IgG anti-PT ELISA met all assay validation parameters within the range considered most relevant for serodiagnosis. This ELISA was developed and analytically validated as a user-friendly kit that can be used in both qualitative and quantitative formats. The technology for producing the kit is transferable to public health laboratories.
The effect of two (2W) versus three (3W) wave patterns of follicular dynamics and concurrent endocrine milieu of follicle‐stimulating hormone (FSH), luteinizing hormone (LH), oestradiol 17‐β (E2) and progesterone (P4) were investigated during one interoestrous interval (IEI) before insemination, on ensuing pregnancy, in 70 lactating Jersey crossbred cows. The findings were evaluated for between [included all (overall) 2W‐O and 3W‐O cows] and within [after separating pregnant (P) and non‐pregnant (NP) cows in 2W and 3W] wave patterns. The propensity of two (58.6%, n = 41) and three (41.4%, n = 29) wave patterns was similar (p = .15). The IEI, shorter by 2.6 days for 2W‐O versus 3W‐O (p < .0009), predicted wave pattern as 100% 2W‐O cows had IEI of ≤21 days, present only in 27.6% 3W‐O cows (p < .0001). The ovulatory follicle persisted for a significantly shorter duration for 3W‐O versus 2W‐O cows. The average FSH, LH, E2 and P4 concentrations during the IEI did not differ for between and within the wave patterns. Pregnancy rate (%) of 58.6 versus 41.4 (p = .15) for 2W‐O versus 3W‐O and 56.1‐P versus 43.9‐NP (p = .44) for within 2W was similar, but tended to differ for within the 3W pattern (69.0‐P versus 31.0‐NP, p = .06). The pregnancy outcome was influenced by the age of ovulatory follicle for between the wave patterns and by follicular count as well as FSH surge concentration for within the wave patterns. A shorter luteal phase reduced the pregnancy outcome, a novel finding of the present study.
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