Accurate detection of α-globin gene copy number variants with two reactions using droplet digital PCR

Bao, Xiuqin; Qin, Danqing; Ma, Junwu; Zhou, Xiangcheng; Wang, Jicheng; Yao, Cuize; Zhang, Liang; Du, Li

doi:10.1080/16078454.2022.2030885

Cited by 5 publications

(2 citation statements)

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“…There are other targeted approaches to the identification of multiple copy number variations (CNVs) in the α-globin genes. [25][26][27][28][29][30] Bao et al, 31 recently found droplet-digital PCR to be 100% accurate and sensitive in the detection of α-globin gene number variants. They recommended this method for screening, followed by another confirmatory technique such as gap-PCR or MLPA.…”

Section: Discussionmentioning

confidence: 99%

Hematological and genetic profiles of persons with co‐inherited heterozygous β‐thalassemia and supernumerary α‐globin genes

Sundaresan

Hira

Chhabra

et al. 2023

European J of Haematology

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Introduction: Thalassemias are common monogenic autosomal recessive hemoglobin disorders. The usually asymptomatic heterozygotes (β-thalassemia traits, βTT) may rarely develop non-transfusion-dependent-thalassemia (NTDT) due to co-inheritance of supernumerary α-globin genes. Literature on phenotypic/genotypic features of these rare combinations is limited. Materials and Methods:We studied the demographic, clinical, and laboratory data from 47 persons with co-inherited βTT + supernumerary α-globin genes. HBB mutations were tested for by ARMS-PCR and/or Sanger sequencing, ααα (anti3.7) /ααα (anti4.2) and deletional α-thalassemia testing by multiplex gap-PCRs, and Xmn1 G γ genotyping by PCR-RFLP. Results:The 47 cases comprised 0.08% of 61 010 hemoglobinopathy screenings during the study period. Mean age was 31.9 ± 14.7 years (range 5.5-83 years), with 57.4% males. Thirty (63.8%) had NTDT-phenotype, 16 (34%) were asymptomatic/ minimally symptomatic, and 1 became transfusion-dependent at the age of 20 years.Anemia/pallor and jaundice were the commonest complaints (76% each); 40% had required blood transfusions. Twenty-one had splenomegaly, 14 had hepatomegaly.Mean hemoglobin was 9.0 ± 1.9 g/dl (range 4.0-13.0). HbA2 was 5.1 ± 0.7% (3.4%-6.3%) and HbF% 4.2 ± 3.2% (0.5%-18.4%). Forty-four (93.6%) had ααα anti3.7 , while 3 (6.4%) had ααα anti4.2 triplications. HBB:c.92+5G>C (47%), HBB:c.27_28insG (14.9%), and HBB:c.47G>A (8.5%) were the commonest β-globin mutations. One case showed HBB:c.-138C>T (β ++ ), while the rest had β 0 or severe-β + mutations. Symptomatic cases had significantly lower hemoglobins and higher HbF% than asymptomatic ones.

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Section: Discussionmentioning

confidence: 99%

Hematological and genetic profiles of persons with co‐inherited heterozygous β‐thalassemia and supernumerary α‐globin genes

Sundaresan

Hira

Chhabra

et al. 2023

European J of Haematology

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“…We used ddPCR to calculate the copy number of HBA1 and HBA2 . ddPCR was performed as previously described [ 8 ]. We used RPP30 to calculate the copy number, which had two copy numbers of RPP30 gene, as the reference gene.…”

Section: Methodsmentioning

confidence: 99%

Identification of four novel large deletions and complex variants in the α-globin locus in Chinese population

et al. 2023

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Background At present, the methods generally used to detect α-thalassemia mutations are confined to detecting common mutations, which may lead to misdiagnosis or missed diagnosis. The single-molecule real-time (SMRT) sequencing enables long-read single-molecule sequencing with high detection accuracy, and long-length DNA chain reads in high-fidelity read mode. This study aimed to identify novel large deletions and complex variants in the α-globin locus in Chinese population. Methods We used SMRT sequencing to detect rare and complex variants in the α-globin locus in four individuals whose hematological data indicated microcytic hypochromic anemia. However, the conventional thalassemia detection result was negative. Multiplex ligation-dependent probe amplification and droplet digital polymerase chain reaction were used to confirm SMRT sequencing results. Results Four novel large deletions were observed ranging from 23 to 81 kb in the α-globin locus. One patient also had a duplication of upstream of HBZ in the deletional region, while another, with a 27.31-kb deletion on chromosome 16 (hg 38), had abnormal hemoglobin Siriraj (Hb Siriraj). Conclusion We first identified the four novel deletions in the α-globin locus using SMRT sequencing. Considering that the conventional methods might lead to misdiagnosis or missed diagnosis, SMRT sequencing proved to be an excellent method to discover rare and complex variants in thalassemia, especially in prenatal diagnosis.

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The -α^3.7III subtype of α⁺-thalassemia was identified in China

et al. 2022

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Accurate detection of α-globin gene copy number variants with two reactions using droplet digital PCR

Cited by 5 publications

References 16 publications

Hematological and genetic profiles of persons with co‐inherited heterozygous β‐thalassemia and supernumerary α‐globin genes

Hematological and genetic profiles of persons with co‐inherited heterozygous β‐thalassemia and supernumerary α‐globin genes

Identification of four novel large deletions and complex variants in the α-globin locus in Chinese population

The -α^3.7III subtype of α⁺-thalassemia was identified in China

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Accurate detection of α-globin gene copy number variants with two reactions using droplet digital PCR

Cited by 5 publications

References 16 publications

Hematological and genetic profiles of persons with co‐inherited heterozygous β‐thalassemia and supernumerary α‐globin genes

Hematological and genetic profiles of persons with co‐inherited heterozygous β‐thalassemia and supernumerary α‐globin genes

Identification of four novel large deletions and complex variants in the α-globin locus in Chinese population

The -α3.7III subtype of α+-thalassemia was identified in China

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The -α^3.7III subtype of α⁺-thalassemia was identified in China