title: [A novel CD36 mutation T538C (Trp180Arg) results in CD36 deficiency and establishment of a genotyping method for the novel mutation based on sequence-specific primer PCR].
abstract: OBJECTIVE
  To explore the molecular basis for a CD36 deficiency individual and distribution of CD36 gene mutation in Guangxi population.
  
  
  METHODS
  A female individual was studied. CD36 phenotype was detected by monoclonal antibody immobilization of platelet antigens assay (MAIPA) and flow cytometry (FCM). The coding regions of the CD36 gene were sequenced. A DNA-based polymerase chain reaction-sequence specific primer (PCR-SSP) assay was used to verify the identified mutation. Cell lines expressing the mutant and wild-type CD36[CD36(MT) and CD36(WT)] were established, with the expression of CD36 determined by Western blotting. The distribution of CD36 gene mutation was investigated among 1010 unrelated individuals with the PCR-SSP assay.
  
  
  RESULTS
  Both MAIPA and FCM assays showed that the patient had type II CD36 deficiency. DNA sequencing showed that she has carried a heterozygous mutation T538C (Trp180Arg) in the exon 6 of CD36. Sequencing of cDNA clone confirmed that there was a nucleotide substitution at position 538 (538T>C). Western blotting also confirmed that the CD36 did not express on the CD36(MT) cell line that expressed the 538C mutant, but did express on the CD36(WT) cell line. The novel CD36 mutation T538C was further verified with 100% concordance of genotyping results by DNA-based PCR-SSP assay and 1010 unrelated individuals. No CD36 538C allele was detected among the 1010 individuals.
  
  
  CONCLUSIONS
  This study has identified a novel CD36 mutation T538C(Trp180Arg)(GenBank: HM217022.1), and established a genotyping method for the novel sequence-specific primer PCR. The novel mutation is rare in Guangxi and can cause type II CD36 deficiency.
authors: Nanning Institute of Transfusion Medicine, Nanning, Guangxi 530007, China; Nanning Blood Center, Nanning, Guangxi 530007, China. Email: guangwu@szonline.net.
authors: Lilan
authors: L
authors: Li
authors: Baoren
authors: B
authors: He
authors: Yan
authors: Y
authors: Zhou
authors: Zhoulin
authors: Z
authors: Zhong
authors: Haiyan
authors: H
authors: Li
authors: Fang
authors: F
authors: Lu
authors: Jinlian
authors: J
authors: Liu
authors: Weidong
authors: W
authors: Shen
authors: Hengcong
authors: H
authors: Li
authors: Lihong
authors: L
authors: Jiang
authors: Guoguang
authors: G
authors: Wu
mesh_terms: D001483:Base Sequence; D001791:Blood Platelet Disorders / Q000235:genetics*; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism; D015153:Blotting, Western; D018955:CD36 Antigens / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000235:genetics; D005091:Exons / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D030342:Genetic Diseases, Inborn / Q000235:genetics*; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D018955:CD36 Antigens; D017931:DNA Primers