Identification of serum proteomic biomarkers for early porcine reproductive and respiratory syndrome (PRRS) infection |
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Authors: | Sem Genini Thomas Paternoster Alessia Costa Sara Botti Mario Vittorio Luini Andrea Caprera Elisabetta Giuffra |
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Institution: | 1. Parco Tecnologico Padano - CERSA, Via Einstein, 26900, Lodi, Italy 5. Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA 2. IASMA-FEM Research and Innovation Centre, Via E. Mach 1, 38010, San Michele a/A, TN, Italy 6. Sandoz Industrial Products S.p.A., Corso Verona 165, 38068, Rovereto, TN, Italy 3. BIOTRACK s.r.l., Via Einstein, 26900, Lodi, Italy 4. Istituto Zooprofilattico Sperimentale della Lombardia e dell??Emilia Romagna, 26900, Lodi, Italy 7. INRA, UMR 1313 de G??n??tique Animale et Biologie Int??grative, Jouy-en-Josas, France
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Abstract: | ABSTRACT: BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) is one of the most significant swine diseases worldwide. Despite its relevance, serum biomarkers associated with early-onset viral infection, when clinical signs are not detectable and the disease is characterized by a weak anti-viral response and persistent infection, have not yet been identified. Surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF MS) is a reproducible, accurate, and simple method for the identification of biomarker proteins related to disease in serum. This work describes the SELDI-TOF MS analyses of sera of 60 PRRSV-positive and 60 PRRSV-negative, as measured by PCR, asymptomatic Large White piglets at weaning. Sera with comparable and low content of hemoglobin (< 4.52 ug/mL) were fractionated in 6 different fractions by anion-exchange chromatography and protein profiles in the mass range 1-200 kDa were obtained with the CM10, IMAC30, and H50 surfaces. RESULTS: A total of 200 significant peaks (p < 0.05) were identified in the initial discovery phase of the study and 47 of them were confirmed in the validation phase. The majority of peaks (42) were up-regulated in PRRSV-positive piglets, while 5 were down-regulated. A panel of 14 discriminatory peaks identified in fraction 1 (pH=9), on the surface CM10, and acquired at low focus mass provided a serum protein profile diagnostic pattern that enabled to discriminate between PRRSV-positive and -negative piglets with a sensitivity and specificity of 77% and 73%, respectively. CONCLUSIONS: SELDI-TOF MS profiling of sera from PRRSV-positive and PRRSV-negative asymptomatic piglets provided a proteomic signature with large scale diagnostic potential for early identification of PRRSV infection in weaning piglets. Furthermore, SELDI-TOF protein markers represent a refined phenotype of PRRSV infection that might be useful for whole genome association studies. |
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