ABSTRACT
OBJECTIVES: The current study aimed at the rapid identification of the copy number of α-globin genes for the diagnosis of α-thalassemia. DESIGN AND METHODS: To identify the copy number of α-globin genes in α-thalassemia, we developed a novel method using a multiplex polymerase chain reaction (PCR) in combination with the CE analysis. RESULTS: The proposed method provides a rapid detection of the common α-globin gene deletions. Sixty-six patients with α-thalassemia and 46 normal controls were included in the present study. The obtained results showed good correlation with those obtained by gap PCR. Moreover, a low amount of maternal cell contamination in the fetus specimen for the prenatal diagnosis of hemoglobin Barts hydrops fetalis as well as the rare multiplicated α-globin genes can be identified using this method. CONCLUSION: This method provides a convenient and efficient tool for the rapid identification of the copy number of α-globin genes in α-thalassemia and the individuals with α-globin gene multiplication.
Subject(s)
Electrophoresis, Capillary/methods , Gene Dosage , alpha-Globins/genetics , alpha-Thalassemia/genetics , Base Sequence , DNA Copy Number Variations/genetics , Female , Gene Deletion , Glycated Hemoglobin/genetics , Hemoglobin A2/genetics , Hemoglobins, Abnormal/genetics , Humans , Hydrops Fetalis/diagnosis , Hydrops Fetalis/genetics , Male , Molecular Sequence Data , Multiplex Polymerase Chain Reaction/methods , Pregnancy , Prenatal Diagnosis/methods , Reproducibility of Results , Sensitivity and Specificity , Sequence Homology, Nucleic Acid , alpha-Thalassemia/diagnosisABSTRACT
BACKGROUND: Reduction of E-cadherin in most common epithelial tumors relates to metastasis, which results from the silence of E-cadherin by CpG methylation. MATERIALS AND METHODS: We examined the E-cadherin expression by immunohistochemical staining and detected methylation by methylation-specific polymerase chain reaction (MSP) in 48 primary oral SCC tissues. RESULTS: The results showed that 41 out of 48 (85.4%) cancerous tissues and 16 out of 48 (33.3%) nearby non-cancerous tissues had CpG methylation on the promoter region of E-cadherin. In these non-cancerous tissues, 2 out of 16 (12.5%) had no methylation change in their paired cancerous part. Immunohistochemical study showed that a decreased expression pattern was found in the tissue which had CpG methylation on the promoter region, but an over expression island or aberrant expression was also frequently found in these cases. CONCLUSION: The methylation of E-cadherin in oral SCC may occur in the precancerous stage and the process is dynamic, which has no relationship with the aberrant expression of E-cadherin protein.
