Unambiguous diagnostic testing for Barth Syndrome (BTHS) is routinely performed bydetermination of the relative amounts and distribution of monolysocardiolipin (MLCL)and cardiolipin (CL) species, confi rmed by TAZ gene sequencing or vice versa. Molecularanalysis of the TAZ gene is a powerful tool for the diagnosis of BTHS but can lead to falsenegative results when mutations are present in regulating or relevant non-codingsequences. Because of this, CL abnormalities in BTHS have been recognised to be animportant biomarker for the diagnosis. Recently we described a novel MALDI-TOF-MS approach for the lipid analysis of intactmitochondria and bacterial cells that does not require extraction and separation steps,leading to a fast and accurate, qualitative and quantitative determination of thecardiolipin levels using a very small amount of sample. Therefore we have used MALDI-MS to develop a new, fast, easy, inexpensive and non-invasive method for the diagnosisof BTHS. The method needs only 1 ml of whole blood; leukocytes are easily isolated by eliminatingerythrocytes with dextran sedimentation followed by hypotonic lysis of residualerythrocytes. Collected leukocytes are then easily analysed by MALDI-MS as intact cells,skipping the lipid extraction procedure.
Monolysocardiolipin/cardiolipin ratio of intact leukocytes as novel tool for the screening of Barth Syndrome
Corcelli Angela;Angelini Roberto;Lobasso Simona;
2015-01-01
Abstract
Unambiguous diagnostic testing for Barth Syndrome (BTHS) is routinely performed bydetermination of the relative amounts and distribution of monolysocardiolipin (MLCL)and cardiolipin (CL) species, confi rmed by TAZ gene sequencing or vice versa. Molecularanalysis of the TAZ gene is a powerful tool for the diagnosis of BTHS but can lead to falsenegative results when mutations are present in regulating or relevant non-codingsequences. Because of this, CL abnormalities in BTHS have been recognised to be animportant biomarker for the diagnosis. Recently we described a novel MALDI-TOF-MS approach for the lipid analysis of intactmitochondria and bacterial cells that does not require extraction and separation steps,leading to a fast and accurate, qualitative and quantitative determination of thecardiolipin levels using a very small amount of sample. Therefore we have used MALDI-MS to develop a new, fast, easy, inexpensive and non-invasive method for the diagnosisof BTHS. The method needs only 1 ml of whole blood; leukocytes are easily isolated by eliminatingerythrocytes with dextran sedimentation followed by hypotonic lysis of residualerythrocytes. Collected leukocytes are then easily analysed by MALDI-MS as intact cells,skipping the lipid extraction procedure.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
