Parra Muñoz, María Dolores

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Parra Muñoz, María Dolores

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Parra Muñoz, María Dolores

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Universidad de Murcia. Departamento de Ciencias Sociosanitarias

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Open Access
Comparación de la 2,2’-ditiodipiridina y el ácido 5,5’-ditiobis-2-nitrobenzoico en la determinación de colinesterasa en sangre entera de perro
(Murcia: Universidad de Murcia, Servicio de publicaciones, 2000) Tecles Vicente, Fernando; Gutiérrez Panizo, Cándido; Martínez Subiela, Silvia; Bolio, M.; Cerón Madrigal, José Joaquín; Parra Muñoz, María Dolores; Facultad de Veterinaria
Embargo
C-reactive protein measurement in canine saliva
(SAGE Publications, 2005-03) Tecles, Fernando; Martínez Subiela, Silvia; Cerón Madrigal, José Joaquín; Parra Muñoz, María Dolores; Ciencias Sociosanitarias
An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 mg/ml and 0.0091 mg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7–9.9% and 8.5–16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R2 5 0.76) and clinical studies (R2 5 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments.
Open Access
Porcine acute phase protein concentrations in different diseases in field conditions
(Wiley, 2006-12) Fuentes, P.; Martínez Subiela, Silvia; Martínez, J.S.; Muñoz, A.; Cerón Madrigal, José Joaquín; Parra Muñoz, María Dolores; Ciencias Sociosanitarias
Five acute phase proteins (APPs) [C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), pig-MAP and albumin] were measured in pigs with naturally occurring infections by porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky’s disease virus (ADV), porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae, and in animals with tail and ear bites, arthritis and other acute inflammatory processes. Healthy specific pathogen-free (SPF) pigs were used as controls. In PRRSV-infected pigs, all APPs with the exception of pig-MAP exhibited significant changes compared with controls. In animals affected with ADV only Hp presented changes of statistical significance, whereas pigs with PCV2 showed marked modifications in all APPs tested. Animals affected with Mycoplasmosis showed concentrations of all positive APPs significantly above levels obtained in SPF pigs, though albumin concentrations did not differ from controls. Finally, all APPs studied showed substantial changes in pigs with acute inflammation. The results indicated that an acute phase response was developed in the different diseases studied, this response being higher in animals with clinical signs and concurrent bacterial processes. Haptoglobin would be the APP that better reflects pathological states; however, to get more complete and valuable information it might be advisable to perform APPs profiles including another APP, such as CRP or SAA.
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Desarrollo de fluoroinmunoensayos en tiempo retardado para determinar proteína C-reactiva y haptoglobina en la especie canina : estudios de validación analítica y clínica / María Dolores Parra Muñoz; directores, José Joaquín Cerón Madrigal y Juan Cabezas Herrera.
(Murcia : Universidad de Murcia, Departamento de Medicina y Cirugía Animal,, 2005) Parra Muñoz, María Dolores
Open Access
Use of time-resolved immunofluorometric assay for determination of canine C-reactive protein concentrations in whole blood
(American Veterinary Medical Association, 2005-01-01) Tuomola, Mika; Cabezas-Herrera, Juan; Cerón Madrigal, José Joaquín; Parra Muñoz, María Dolores; Ciencias Sociosanitarias
Objective: To develop and validate a time-resolved immunofluorometric assay (TR-IFMA) for measurement of C-reactive protein (CRP) in canine whole blood. Animals: 12 healthy dogs and 35 dogs with inflammatory processes. Procedure: CRP was isolated from acute-phase serum by affinity chromatography and used as a standard for calibration. Analytic and functional limit of detection and intra-assay and interassay precision were calculated. Accuracy was evaluated by recovery assays and by comparison with results of a commercial ELISA. Correlation between CRP concentrations in whole blood and corresponding plasma fractions was tested by use of TR-IFMA. Stability of blood samples at 4 degrees C was assessed during a 1-month period, and effects of anticoagulants were evaluated. Measurements of CRP in blood samples from 12 healthy dogs were compared with those of 35 dogs with inflammatory diseases. Results: Analytic and functional limits of detection were 0.53 and 3.26 microg/mL, respectively. Intra-assay and interassay coefficients of variation varied between 2.1% to 8.9% and 8.0% to 12.3%, respectively. Mean recoveries of added CRP were 104% and 114%. Measurements of CRP by use of TR-IFMA and ELISA were highly correlated (R2 = 0.97). Measurements of CRP in whole blood and in corresponding plasma fractions by use of TR-IFMA were also highly correlated (R2 = 0.97). Neither storage nor use of anticoagulants disturbed measurement of CRP concentrations in whole blood. Concentrations of CRP in whole blood of dogs with inflammation were significantly higher than in healthy dogs. Conclusions and clinical relevance: Determination of CRP concentrations in whole blood may provide a diagnostic test for inflammation in dogs.
Open Access
Detección de proteínas de fase aguda en cerdos afectados clínicamente por el síndrome de adelgazamiento multisistémico post-destete
(Murcia: Universidad de Murcia, Servicio de Publicaciones, 2005) Pallarés Martínez, Francisco José; Gómez Cabrera, Serafín; Cerón Madrigal, José Joaquín; Ramis Vidal, Manuel Guillermo; Martínez Subiela, Silvia; Tecles Vicente, Fernando; Martínez, J. S.; Gómez Sánchez, Miguel Ángel; Bernabé Salazar, Antonio; Muñoz Luna, Antonio; Seva Alcaraz, Juan; Parra Muñoz, María Dolores; Facultad de Veterinaria
Open Access
Development of a time-resolved fluorometry based immunoassay for the determination of canine haptoglobin in various body fluids
(INRA / EDP Sciences, 2005) Väisänen, Ville; Cerón Madrigal, José Joaquín; Parra Muñoz, María Dolores; Ciencias Sociosanitarias
A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of haptoglobin (Hp) in canine serum. Haptoglobin was purified from canine acute phase serum by ammonium sulphate precipitation followed by gel filtration. This isolated dog Hp was used as the standard to calibrate the assay. Intra- and inter-assay coefficients of variation of the assay were, respectively, 5.7% and 16.6% at 0.51 mg/mL, 2.4% and 10.6% at 2.1 mg/mL and 10.5% and 11.9% at 32.5 mg/mL. The dilution of serum samples with high Hp concentrations resulted in linear regression equations with R2 of 0.99 and 0.97. A high correlation was found in serum Hp measurements by TR-IFMA and a commercial assay based on peroxidase activity of haemoglobin bound to haptoglobin (R2 = 0.96). The limit of detection for the TR-IFMA method was 0.002 µg/mL. The addition of fresh haemolysate to serum samples did not affect the haptoglobin concentration (P = 0.694). Statistical differences (P < 0.003) were found between healthy dogs and dogs with different pathological processes. In whole blood, Hp concentrations were much lower than in serum but closely related (R2 = 0.84) whereas saliva Hp concentrations were poorly related with serum concentrations (R2 =0.53). However, the concentration of Hp in saliva was significantly (P < 0.039) higher in dogs with pathological processes compared to healthy dogs. The assay sensitivity was adequate to also be applied to whole blood and saliva specimens.
Embargo
Dual-label time-resolved fluoroimmunoassay for simultaneous quantification of haptoglobin and C-reactive protein in meat juice from pigs
(Canadian Veterinary Medical Association, 2012-04-02) Gutiérrez, Ana M.; Cerón Madrigal, José Joaquín; Marsilla, Blas A.; Martínez Subiela, Silvia; Parra Muñoz, María Dolores; Ciencias Sociosanitarias
A new method was developed to simultaneously measure 2 acute-phase proteins (APPs) by time-resolved immunofluorometry. The assay, based on double-label quantification of haptoglobin (Hp) and C-reactive protein (CRP) in meat juice samples from pigs, was constructed by use of a combination of europium and samarium chelate lanthanides as labels. Meat juice samples from 154 pigs were used for analytic and clinical validation of the assay through determination of precision, accuracy, limit of detection, and quantification. The analytic performance of the assay was satisfactory, with good intra-assay and interassay precision and accuracy. The levels of Hp and CRP were increased in the meat juice samples of diseased animals compared with healthy ones. According to the results, higher sensitivity could be achieved if the cut-off values of both proteins were taken into account for clinical relevance rather than used individually. Since the dual assay saved both time and sample, it could be used as a rapid and sensitive screening test in porcine production.
Embargo
C-reactive protein measurements in meat juice of pigs
(Elsevier, 2008-04-15) Gutiérrez, A.M.; Martínez Subiela, Silvia; Montes, A.; Cerón Madrigal, José Joaquín; Parra Muñoz, María Dolores; Ciencias Sociosanitarias
A time-resolved immunofluorometric assay was evaluated for measurement of C-reactive protein in meat juice from diaphragmatic muscle collected from slaughtered pigs. Analytical and clinical validation of the method was performed by using meat juice samples, obtained by freezing and thawing muscle pieces. The intra- and inter-assay coefficients of variation ranged from 2.2–5.8% to 7.9–14.3%, respectively. The limit of detection was 0.00038 mg/ml. The method measured the CRP concentrations in a linear manner with a good accuracy (r = 0.99). CRP concentrations in serum were highly correlated with those in diaphragmatic meat juice (r = 0.90; p < 0.001). CRP concentrations were significantly higher in clinically affected pigs compared to non-diseased pigs. The assay described here provides a sensitive method for measuring CRP concentrations in meat juice, which can represent a suitable alternative to serum or blood samples and simplifies the process of sampling collection at slaughter.
Open Access
Concentrations of C-reactive protein in effusions in dogs
(Wiley, British Veterinary Association, 2006-06) Papasouliotis, K.; Cerón Madrigal, José Joaquín; Parra Muñoz, María Dolores; Ciencias Sociosanitarias
The concentration of C-reactive protein (CRP) was measured in effusions from 50 dogs to assess the potential for measuring this protein to differentiate body cavity fluids. The effusions were classified as either transudates, modified transudates or exudates according to their total protein concentration, total nucleated cell count, cytological findings and aetiology, and the concentration of CRP was determined by a time-resolved immunofluorometric assay. There were significant differences between the concentrations of CRP in the three types of effusion; the highest concentrations were observed in the exudates (4·47 to 54·59 µg/ml), the lowest were in the transudates (0·0094 to 7·87 µg/ml), and the modified transudates contained intermediate concentrations of CRP (0·045 to 10·78 µg/ml). A cut-off value of 4 µg/ml had a sensitivity of 100 per cent and a specificity of 94·4 per cent for differentiating transudates from exudates, and a cut-off value of 11 µg/ml had a sensitivity of 88·2 per cent and a specificity of 100 per cent for distinguishing modified transudates from exudates. However, a cut-off value of 1 µg/ml had a lower sensitivity (80 per cent) and an unacceptably low specificity (66·7 per cent) for differentiating transudates from modified transudates.