A More Universal Approach to Comprehensive Analysis of Thalassemia Alleles (CATSA)

Liang, Qiaowei; Gu, Wanqian; Chen, Ping; Li, Yuezhen; Li, Yanqiu; Tian, Minglei; Zhou, Qiang; Qi, Hongbo; Zhang, Yuhong; He, Jun; Li, Qing; Tang, Lingfang; Tang, Juan; Teng, Yuanwen; Zhou, Yulin; Huang, Sheng‐Wen; Lu, Zongjie; Xu, Mengnan; Hou, Wei; Huang, Ting; Li, Youqiong; Li, Rong; Hu, Lanping; Li, Shaoying; Guo, Qiwei; Zhuo, Zhaozhen; Mou, Yan; Cram, David S.; Wu, Lingqian

doi:10.1016/j.jmoldx.2021.06.008

Cited by 65 publications

(63 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Twelve hospitals in southern China assessed a comprehensive analysis of thalassemia alleles (CATSA) for identifying both α and β thalassemia genetic carrier status by third-generation sequencing (TGS). Compared with standard thalassemia variant PCR panel testing, TCS can detected 33 more positive variants, and found that the traditional PCR detection technology had 1 false negative and 8 false positive result [ 6 ]. The present study used the SMRT and conventional technologies to test the thalassemia gene in the thalassemia screening positive population in this area.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, due to the high homology between HBA2 and HBA1 genes, the short-read NGS method cannot distinguish HBA2 and HBA1 effectively [ 3 , 4 ]. With the advantage of long-molecule sequencing, PacBio real-time sequencing technology (SMRT) had been used for comprehensive and precious thalassemia test [ 5 , 6 ]. In this study, SMRT technology and conventional methods were performed for 434 suspected carriers of thalassemia to simultaneously detect deletion and non-deletion variants of α-thalassemia and β-thalassemia.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The value of single-molecule real-time technology in the diagnosis of rare thalassemia variants and analysis of phenotype–genotype correlation

Luo

Zeng²,

Tang³

et al. 2021

J Hum Genet

View full text Add to dashboard Cite

To compare single-molecule real-time technology (SMRT) and conventional genetic diagnostic technology of rare types of thalassemia mutations, and to analyze the molecular characteristics and phenotypes of rare thalassemia gene variants, we used 434 cases with positive hematology screening as the cohort, then used SMRT technology and conventional gene diagnosis technology [(Gap-PCR, multiple ligation probe amplification technology (MLPA), PCR-reverse dot blot (RDB)] for thalassemia gene screening. Among the 434 enrolled cases, conventional technology identified 318 patients with variants (73.27%) and 116 patients without variants (26.73%), SMRT identified 361 patients with variants (83.18%), and 73 patients without variants (16.82%). The positive detection rate of SMRT was 9.91% higher than conventional technology. Combination of the two methods identified 485 positive alleles among 49 types of variant. The genotypes of 354 cases were concordant between the two methods, while 80 cases were discordant. Among the 80 cases, 76 cases had variants only identified in SMRT method, 3 cases had variants only identified in conventional method, and 1 false positive result by the traditional PCR detection technology. Except the three variants in HS40 and HBG1-HBG2 loci, which was beyond the design of SMRT method in this study, all the other discordant variants identified by SMRT were validated by further Sanger sequencing or MLPA. The hematological phenotypic parameters of 80 discordant cases were also analyzed. SMRT technology increased the positive detection rate of thalassemia genes, and detected rare thalassemia cases with variable phenotypes, which had great significance for clinical thalassemia gene screening.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The value of single-molecule real-time technology in the diagnosis of rare thalassemia variants and analysis of phenotype–genotype correlation

Luo

Zeng²,

Tang³

et al. 2021

J Hum Genet

View full text Add to dashboard Cite

show abstract

“…Experiments were performed as previously described 14 . Briefly, genomic DNA was amplified by PCR with primers targeting the majority of known structural variations, SNVs and indels in the HBA1 , HBA2 and HBB genes.…”

Section: Methodsmentioning

confidence: 99%

“…However, for rare variants not located in regular regions or variants in homologous regions of HBA1 and HBA2 , conventional methods and short-read based NGS methods may lead to missed diagnosis or even misdiagnosis. Currently, a methodology based on third-generation sequencing (TGS) named Comprehensive Analysis of ThalaSsaemia Alleles (CATSA) was developed and validated for comprehensive thalassemia screening 14 , 15 . Based on long-range PCR and long-molecule sequencing on the PacBio Sequel II platform, the CATSA method is adequate to cover the entire gene region and enable the diagnosis of common and rare types of thalassemia variants.…”

Section: Introductionmentioning

confidence: 99%

Analysis of rare thalassemia genetic variants based on third-generation sequencing

Cheng

Zhang

Ren

et al. 2022

Sci Rep

View full text Add to dashboard Cite

Thalassemia is a group of common hereditary anemias that cause significant morbidity and mortality worldwide. However, precisely diagnosing thalassemia, especially rare thalassemia variants, is still challenging. Long-range PCR and long-molecule sequencing on the PacBio Sequel II platform utilized in this study could cover the entire HBA1, HBA2 and HBB genes, enabling the diagnosis of most of the common and rare types of thalassemia variants. In this study, 100 cases of suspected thalassemia were subjected to traditional thalassemia testing and third-generation sequencing for thalassemia genetic diagnosis. Compared with traditional diagnostic methods, an additional 10 cases of rare clinically significant variants, including 3 cases of structure variants and 7 cases of single nucleotide variations (SNVs) were identified, of which a case with − α3.7 subtype III (− α3.7III) was first identified and validated in the Chinese population. Other rare variants of 11.1 kb deletions (− 11.1/αα), triplicate α-globin genes (aaa3.7/αα) and rare SNVs have also been thoroughly detected. The results showed that rare thalassemia variants are not rare but have been misdiagnosed by conventional methods. The results further validated third-generation sequencing as a promising method for rare thalassemia genetic testing.

show abstract

“…Recently, there are many diagnosis methods for thalassemia screening [7,8], for example, routine blood analysis by mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV), erythrocyte osmotic fragility test (EOFT), serum iron test (SIT), Hb electrophoresis measurement (HEM), isoelectric focusing (IEF), liquid chromatography (LC), and next-generation sequencing (NGS). Among these methods, MCH and MCV are commonly used for thalassemia screening; however, these two methods are not specific enough.…”

Section: Introductionmentioning

confidence: 99%

Wooden-Tip Electrospray Mass Spectrometry Characterization of Human Hemoglobin in Whole Blood Sample for Thalassemia Screening: A Pilot Study

Huang

Zou

et al. 2022

Molecules

Self Cite

View full text Add to dashboard Cite

Traditional analytical methods for thalassemia screening are needed to process complicated and time-consuming sample pretreatment. In recent decades, ambient mass spectrometry (MS) approaches have been proven to be an effective analytical strategy for direct sample analysis. In this work, we applied ambient MS with wooden-tip electrospray ionization (WT-ESI) for the direct analysis of raw human blood samples that were pre-identified by gene detection. A total of 319 whole blood samples were investigated in this work, including 100 α-thalassemia carriers, 67 β-thalassemia carriers, and 152 control healthy samples. Only one microliter of raw blood sample was directly loaded onto the surface of the wooden tip, and then five microliters of organic solvent and a high voltage of +3.0 kV were applied onto the wooden tip to generate spray ionization. Multiply charged ions of human hemoglobin (Hb) were directly observed by WT-ESI-MS from raw blood samples. The signal ratios of Hb chains were used to characterize two main types of thalassemia (α and β types) and healthy control blood samples. Our results suggested that the ratios of charged ions to Hb chains being at +13 would be an indicator for β-thalassemia screening.

show abstract

A More Universal Approach to Comprehensive Analysis of Thalassemia Alleles (CATSA)

Cited by 65 publications

References 30 publications

The value of single-molecule real-time technology in the diagnosis of rare thalassemia variants and analysis of phenotype–genotype correlation

The value of single-molecule real-time technology in the diagnosis of rare thalassemia variants and analysis of phenotype–genotype correlation

Analysis of rare thalassemia genetic variants based on third-generation sequencing

Wooden-Tip Electrospray Mass Spectrometry Characterization of Human Hemoglobin in Whole Blood Sample for Thalassemia Screening: A Pilot Study

Contact Info

Product

Resources

About