Validation and development of an immunonephelometric assay for the determination of alpha-1 antitrypsin levels in dried blood spots from patients with COPD

• Autor(a) principal: Zillmer,Laura Russo
• Data de Publicação: 2013
• Outros Autores: Russo,Rodrigo, Manzano,Beatriz Martins, Ivanaga,Ivan, Nascimento,Oliver Augusto, Souza,Altay Alves Lino de, Santos Junior,Gildo, Rodriguez,Francisco, Miravitlles,Marc, Jardim,Jose Roberto
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Jornal Brasileiro de Pneumologia (Online)
• Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1806-37132013000500547
• topic: alpha 1-antitrypsin deficiency; Nephelometry and turbidimetry; Pulmonary disease, chronic obstructive

OBJECTIVE: To validate and develop an immunonephelometric assay for the determination of alpha-1 antitrypsin (AAT) levels in dried blood spots from COPD patients in Brazil. METHODS: We determined AAT levels in serum samples and dried blood spots from 192 COPD patients. For the preparation of dried blood spots, a disk (diameter, 6 mm) was placed into a tube, eluted with 200 µL of PBS, and stored overnight at 4ºC. All of the samples were analyzed by immunonephelometry in duplicate. We used the bootstrap resampling method in order to determine a cut-off point for AAT levels in dried blood spots. RESULTS: The correlation coefficient between the AAT levels in serum samples and those in dried blood spots was r = 0.45. For dried blood spots, the cut-off value was 2.02 mg/dL (97% CI: 1.45-2.64 mg/dL), with a sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 95.7%, 27.2%, and 100%, respectively. CONCLUSIONS: This method for the determination of AAT levels in dried blood spots appears to be a reliable screening tool for patients with AAT deficiency.