Simple method for α 1 ‐antitrypsin deficiency screening by use of dried blood spot specimens

The use of dried blood spot (DBS) specimens in quantitative α 1 ‐antitrypsin (α 1 ‐AT) detection or genetic analysis is limited because protein levels in the samples are low and they contain components that can interfere with polymerase chain reaction amplification. A methodological adaptation was developed to overcome these drawbacks which is discussed here. The study population consisted of 200 healthy volunteers and 300 patients with chronic obstructive pulmonary disease (COPD). DBS specimens were tested for α 1 ‐AT concentration using a modified nephelometric assay and phenotyped with an isoelectric focusing method. Genetic diagnosis was established by deoxyribonucleic acid sequencing using a simple purification procedure to remove contaminants. The nephelometric method showed a detection limit of 0.284 mg·dL ‐1 , corresponding to a serum concentration of 13 mg·dL ‐1 . The correlation coefficient between α 1 ‐AT concentrations in DBS versus serum samples was R 2 =0.8674 (p<0.0001). All 200 healthy individuals had DBS α 1 ‐AT concentrations >1.9 mg·dL ‐1 , corresponding to 114 mg·dL ‐1 in serum samples. One hundred and twenty‐five COPD patients (42%) showed α 1 ‐AT values <1.8 mg·dL ‐1 . Twenty patients with the PIZ phenotype had α 1 ‐AT values lower than 0.64 mg·dL ‐1 . On the basis of genotyping, one COPD patient was classified as heterozygous (PIMMheerlen). Selective elution of contaminants resulted in optimal α 1 1‐antitrypsin genotyping. Because of its sensitivity and excellent correlation with the standard method, the dried blood spot quantitative assay is a reliable tool for routine measurement of α 1 ‐antitrypsin.