Q fever remains a One Health problem, posing a zoonotic threat and causing significant economic losses to the livestock industry. The advancement of detection tools is critical to the effective control of infection. In humans, laboratory investigations depend largely on the immunofluorescence assay, considered the gold standard. In contrast, serologic tools routinely used for veterinary screening have several gaps, resulting in interpretations that are frequently misleading. We investigated the potential application of recombinant Ybgf antigen (r-Ybgf), a periplasmic protein described as one of the most immunodominant antigens in humans, in an indirect ELISA. Following successful expression in the prokaryotic system and the preliminary evaluation of immunoreactivity in western blot, we used r-Ybgf to develop an in-house ELISA using serum samples from sheep, goats, and cattle, which were tested in parallel with an Idexx ELISA kit. The results obtained with the 2 tests were compared, and r-Ybgf performed favorably, with 81.8% sensitivity and 90.1% specificity and substantial agreement, as revealed by receiver operating characteristic analysis. Moreover, we evaluated the serologic response against phase I (PhI) and phase II (PhII) antigens, and r-Ybgf antigen induced by vaccination, using phase-specific ELISAs. The dynamics of antibody response showed a significant increase in reactivity against PhI and PhII, but not against r-Ybgf, antigens. This property may be very useful given the absence of a protocol for the differentiation of infected from vaccinated animals.

Characterization of recombinant Ybgf protein for the detection of Coxiella antibodies in ruminants

Colitti B.;Rosati S.;
2022-01-01

Abstract

Q fever remains a One Health problem, posing a zoonotic threat and causing significant economic losses to the livestock industry. The advancement of detection tools is critical to the effective control of infection. In humans, laboratory investigations depend largely on the immunofluorescence assay, considered the gold standard. In contrast, serologic tools routinely used for veterinary screening have several gaps, resulting in interpretations that are frequently misleading. We investigated the potential application of recombinant Ybgf antigen (r-Ybgf), a periplasmic protein described as one of the most immunodominant antigens in humans, in an indirect ELISA. Following successful expression in the prokaryotic system and the preliminary evaluation of immunoreactivity in western blot, we used r-Ybgf to develop an in-house ELISA using serum samples from sheep, goats, and cattle, which were tested in parallel with an Idexx ELISA kit. The results obtained with the 2 tests were compared, and r-Ybgf performed favorably, with 81.8% sensitivity and 90.1% specificity and substantial agreement, as revealed by receiver operating characteristic analysis. Moreover, we evaluated the serologic response against phase I (PhI) and phase II (PhII) antigens, and r-Ybgf antigen induced by vaccination, using phase-specific ELISAs. The dynamics of antibody response showed a significant increase in reactivity against PhI and PhII, but not against r-Ybgf, antigens. This property may be very useful given the absence of a protocol for the differentiation of infected from vaccinated animals.
2022
34
4
646
653
coxiellosis; Q fever; recombinant antigen; recombinant ELISA; Animals; Cattle; Coxiella; Enzyme-Linked Immunosorbent Assay; Goats; Recombinant Proteins; Ruminants; Sensitivity and Specificity; Sheep; Coxiella burnetii; Goat Diseases; Q Fever
Ferrara G.; Colitti B.; Pagnini U.; Iovane G.; Rosati S.; Montagnaro S.
File in questo prodotto:
File Dimensione Formato  
Characterization of recombinant Ybgf protein for the detection of Coxiella antibodies in ruminants Enhanced Reader.pdf.zip

Accesso aperto

Dimensione 6.79 MB
Formato Zip File
6.79 MB Zip File Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2318/1881246
Citazioni
  • ???jsp.display-item.citation.pmc??? 1
  • Scopus 6
  • ???jsp.display-item.citation.isi??? 6
social impact