Molecular analysis of patients with type III Bartter syndrome: picking up large heterozygous deletions with semiquantitative PCR.
Type III Bartter syndrome (BS) (OMIM607364) is caused by mutations in the basolateral chloride channel CIC-Kb gene (CLCNKB). The CLCNKB gene is sometimes reported as having a large deletion mutation, but all cases reported previously were large homozygous deletions and a large heterozygous deletion is impossible to detect by direct sequencing. This report concerns a genetic analysis of five Japanese patients with type III BS. To identify the mutations, we used polymerase chain reaction (PCR) and direct sequencing. To detect large heterozygous deletion mutations of the CLCNKB gene, we conducted semiquantitative PCR amplification using capillary electrophoresis. The result was that four mutations were identified, comprising one novel 2-bp deletion mutation, an entire heterozygous deletion, and a heterozygous deletion mutation of exons 1 and 2. The nonsense mutation W610X was detected in all patients, and this mutation is likely to constitute a founder effect in Japan. Capillary electrophoresis is a new method and extremely useful for detecting large heterozygous deletions, and should be used to examine type III BS cases in whom only a heterozygous mutation has been detected by direct sequencing. This is the first report to identify large heterozygous deletion mutations in the CLCNKB gene in patients with type III BS.
Nozu
Kandai
K
Department of Pediatrics, Kobe University Graduate School of Medicine, Kobe, Hyogo, 6500017, Japan. nozu@med.kobe-u.ac.jp
Fu
Xue Jun
XJ
Nakanishi
Koichi
K
Yoshikawa
Norishige
N
Kaito
Hiroshi
H
Kanda
Kyoko
K
Krol
Rafal Przybyslaw
RP
Miyashita
Ritsuko
R
Kamitsuji
Hidekazu
H
Kanda
Shoichiro
S
Hayashi
Yoshiki
Y
Satomura
Kenichi
K
Shimizu
Nobuhiko
N
Iijima
Kazumoto
K
Matsuo
Masafumi
M
D000293:Adolescent; D000328:Adult; D001477:Bartter Syndrome / Q000235:genetics*; D002675:Child, Preschool; D018118:Chloride Channels / Q000235:genetics; D004252:DNA Mutational Analysis; D005260:Female; D005819:Genetic Markers; D006239:Haplotypes; D006579:Heterozygote; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D017384:Sequence Deletion
C412733:CLCNKB protein, human; D018118:Chloride Channels; D005819:Genetic Markers