One single-tube PCR assay to detect both CD17 (A>T) and IVS-II nt-654 (C>T) homozygous mutations of β-thalassemia

One single-tube PCR assay to detect both CD17 (A>T) and IVS-II nt-654 (C>T) homozygous mutations of β-thalassemia

Aim: Several methods can be used to detect gene mutations associated with β-thalassemia, but it is difficult to achieve reliable and reproducible results. In this study, we introduce a new method, a single-tube multiplex polymerase chain reaction (PCR) assay, to detect both CD17 (A>T) and IVS-II nt-654 (C>T) homozygous mutations. Materials and methods: This new method designs specific primers to diagnose homozygous mutations and normal controls. Results: After PCR amplification, homozygous mutations produce different fragments from normal controls. Conclusion: This study represents an important step towards the development of a novel protocol to diagnose β-thalassemia and other diseases that target numerous mutations.

___

  • 1. Bunn HF, Forger BG. Hemoglobin: Molecular, Genetic and Clinical Aspects, 1st ed. Philadelphia: WB Saunders; 1986.
  • 2. Batebi A, Pourreza A, Esmailian R. Discrimination of betathalassemia minor and iron deficiency anemia by screening test for red blood cell indices. Turk J Med Sci 2012; 42: 275–80.
  • 3. Orkin SH, Kazazian HH Jr. The mutation and polymorphism of the human beta-globin gene and its surrounding DNA. Annu Rev Genet 1984; 18: 131–71.
  • 4. Antonarakis SE, Irkin SH, Cheng TC, Scott AF, Sexton JP, Trusko SP et al. β-Thalassemia in American Blacks: novel mutations in the “TATA” box and an acceptor splice site. Proc Natl Acad Sci USA 1984; 81: 1154–8.
  • 5. Kazazian HH Jr, Orkin SH, Antonarakis SE, Sexton JP, Boehm CD, Goff SC et al. Molecular characterization of seven betathalassemia mutations in Asian Indians. EMBO J 1984; 3: 593– 6.
  • 6. Cheng TC, Orkin SH, Antonarakis SE, Potter MJ, Sexton JP, Markham AF et al. β-Thalassemia in Chinese: use of in vivo RNA analysis and oligonucleotide hybridization in systematic characterization of molecular defects. Proc Natl Acad Sci USA 1984; 81: 2821–5.
  • 7. Chan V, Chan TK, Chebab FF, Todd D. Distribution of betathalassemia mutations in south China and their association with haplotypes. Am J Hum Genet 1987; 41: 678–85.
  • 8. Xu XM, Zhou YQ, Luo GX, Liao C, Zhou M, Chen PY et al. The prevalence and spectrum of alpha and beta thalassaemia in Guangdong Province: implications for the future health burden and population screening. J Clin Pathol 2004; 57: 517–22.
  • 9. Lin M, Zhu JJ, Wang Q, Xie LX, Lu M, Wang JL et al. Development and evaluation of a reverse dot blot assay for the simultaneous detection of common alpha and beta thalassemia in Chinese. Blood Cells Mol Dis 2012; 48: 86–90.
  • 10. Cao A, Galanello R. Beta-thalassemia. Genet Med 2010; 12: 61–76.
  • 11. Vrettou C, Traeger-Synodinos J, Tzetis M, Malamis G, Kanavakis E. Rapid screening of multiple beta-globin gene mutations by real-time PCR on the LightCycler: application to carrier screening and prenatal diagnosis of thalassemia syndromes. Clin Chem 2003; 49: 769–76.
  • 12. Ye BC, Zhang Z, Lei Z. Molecular analysis of alpha/betathalassemia in a southern Chinese population. Genet Test 2007; 11: 75–83.
  • 13. Clark BE, Thein SL. Molecular diagnosis of haemoglobin disorders. Clin Lab Haematol 2004; 26: 159–76.
  • 14. Xiong F, Sun M, Zhang X, Cai R, Zhou Y, Lou J et al. Molecular epidemiological survey of haemoglobinopathies in the Guangxi Zhuang Autonomous Region of southern China. Clin Genet 2010; 78: 139–48.
  • 15. Xu XM, Zhou YQ, Luo GX, Liao C, Zhou M, Chen PY et al. The prevalence and spectrum of α- and β-thalassemia in Guangdong Province: implications for the future health burden and population screening. J Clin Pathol 2004; 57: 517–22.
Turkish Journal of Medical Sciences-Cover
  • ISSN: 1300-0144
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

Helicobacter pylori infection and relationship with gastric epithelial cell proliferation and apoptosis

Zühal GÜCİN, Tuğba ÇAKMAK, Nizamettin BAYYURT, Barik SALİH

Protective effect of vardenafil on ischemia–reperfusion injury in rat ovary

Hatice BAŞ, Özlem KARA, Mustafa KARA, Dilek PANDIR

Contribution of spirometry to early diagnosis of chronic obstructive pulmonary disease in primary health care centers

Aysel ERDOĞAN, Elif Yilmazel UÇAR, Ömer ARAZ, Leyla SAĞLAM, Nihal Arzu MİRİCİ

Prevalence of type 1 diabetes mellitus in school children 6 18 years old in Diyarbakır, Southeastern Anatolian Region of Turkey

Hüseyin DEMİRBİLEK, Mehmet Nuri ÖZBEK

Serum leptin levels and their proportional relationship with pro- and antiinflammatory mediators in aggressive periodontitis: preliminary report

Zuhal YETKİN AY, Recep SÜTÇÜ, Havva KOÇAK, Fatma Yeşim KIRZIOĞLU

A potential association between the number of CA repeats in the promoter region of the ADAMTS9 gene with lymphatic metastasis of breast cancer

Mikdat BOZER, Fatma AŞIK, Muradiye ACAR, Hacer HALTAŞ, Sibel YENİDÜNYA, Metin CANBAL, Vehap TOPÇU, Muhammet Ramazan YİĞİTOĞLU, Mehmet GÜNDÜZ, Esra GÜNDÜZ, Satoshi HİROHATA, Kadir DEMİRCAN

Posttonsillectomy pain relief and epithelialization with honey

Pathma LETCHUMANAN, Raman RAJAGOPALAN, Mohd Yusoff KAMARUDDIN

Prevalence of type 1 diabetes mellitus in school children 6–18 years old in Diyarbakır, Southeastern Anatolian Region of Turkey

Hüseyin DEMİRBİLEK, Mehmet Nuri ÖZBEK

The effect of maternal–fetal attachment education on maternal mental health

Elieh ABASI, Mahin TAFAZZOLI, Habibolah ESMAILY, Hossain HASANABADI

Comparison of sternal intramedullary bleeding prevention strategies in cardiac surgery

Murat TAVLAŞOĞLU, Ahmet Barış DURUKAN, Mustafa KÜRKLÜOĞLU, Adem GÜLER