Candidate gene analysis using genomic quantitative PCR: identification of ADAMTS13 large deletions in two patients with Upshaw‐Schulman syndrome. Issue 3 (14th January 2014)

• Record Type: Journal Article
• Authors: Eura, Yuka; Kokame, Koichi; Takafuta, Toshiro; Tanaka, Ryojiro; Kobayashi, Hikaru; Ishida, Fumihiro; Hisanaga, Shuichi; Matsumoto, Masanori; Fujimura, Yoshihiro; Miyata, Toshiyuki
• Is Part Of: Molecular genetics & genomic medicine. Volume 2:Issue 3(2014:May)
• Page Start: 240
• Page End: 244
• Publication Date: 2014-01-14
• Subjects: Medical genetics -- Periodicals; Genomics -- Periodicals; 616.042
• DOI: 10.1002/mgg3.64
• Languages: English
• ISSNs: 2324-9269

Abstract

<abstract abstract-type="main" id="mgg364-abs-0001"> <title>Abstract</title> <p>Direct sequencing is a popular method to discover mutations in candidate genes responsible for hereditary diseases. A certain type of mutation, however, can be missed by the method. Here, we report a comprehensive genomic quantitative polymerase chain reaction (qPCR) to complement the weakness of direct sequencing. Upshaw‐Schulman syndrome (USS) is a recessively inherited disease associated with severe deficiency of plasma ADAMTS13 activity. We previously analyzed <italic>ADAMTS13</italic> in 47 USS patients using direct sequencing, and 44 of them had either homozygous or compound heterozygous mutations. Then, we sought to reveal more extensive defects of <italic>ADAMTS13</italic> in the remaining three patients. We quantified copy numbers of each <italic>ADAMTS13</italic> exon in the patients by using genomic qPCR. Each primer pair was designed to contain at least one of the two primers used in direct sequencing, to avoid missing any exonic deletions. The qPCR demonstrated heterozygous loss of exons 7 and 8 in one patient and exon 27 in the other, and further analysis revealed c.746_987+373del1782 and c.3751_3892+587del729, respectively. Genomic qPCR provides an effective method for identifying extensive defects of the target genes.</p> </abstract>