Using DNA testing for the precise, definite, and low-cost diagnosis of sickle cell disease and other Haemoglobinopathies: findings from Tanzania

Abstract Background Sickle cell disease (SCD) is an important cause of under-five mortality. Tanzania is the 5th country in the world with the highest births prevalence of SCD individuals. Significant advances in the neonatal diagnosis of SCD using rapid point-of-care testing have been made. However genetic confirmation is still required for positive cases, in uncertain cases, in multiply transfused patients, to resolve compound heterozygosity (Hb S/ β0 Thal or Hb S/ β+ thal) not uncommon in the coastal regions of East Africa and increasingly also for pre-marital counselling and potentially for future curative approaches such as gene therapy. The currently available DNA tests are prohibitively expensive. Here, we describe an easy-to-use, affordable and accurate β-globin sequencing approach that can be easily integrated within existing NBS for SCD and other haemoglobinopathies especially in Low- and Middle-income Countries. Aim To evaluate an affordable DNA technology for the diagnosis of Sickle cell disease and other haemoglobinopathies in a resource-limited setting. Methods Laboratory-based validation study was conducted by Muhimbili University of Health and Allied Sciences and the University of Oxford involving sequencing of the entire β -haemoglobin locus using the Oxford Nanopore MinION platform. A total number of 36 Dried blood spots and whole blood samples were subjected to conventional protein-based methods (isoelectric focusing, HPLC), and/or sequenced by the Sanger method as comparators. Results Sequencing results for SCD using the MinION were 100% concordant with those from the Sanger method. In addition, the long-read DNA sequencing method enabled the resolution of cases with unusual phenotypes which make up 1% of all children in Tanzania. The cost is £11/ sample for consumables, which is cheaper compared to other sequencing platforms. Conclusions This is the first report of a comprehensive single DNA assay as a definitive diagnostic test for SCD and other haemoglobinopathies. The test is fast, precise, accurate and affordable.

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HemoTypeSC, a low-cost point-of-care testing device for sickle cell disease: Promises and challenges

Blood Cells Molecules and Diseases ◽

10.1016/j.bcmd.2019.01.007 ◽

2019 ◽

Vol 78 ◽

pp. 22-28 ◽

Cited By ~ 8

Author(s):

Obiageli Nnodu ◽

Hezekiah Isa ◽

Maxwell Nwegbu ◽

Chinatu Ohiaeri ◽

Samuel Adegoke ◽

...

Keyword(s):

Sickle Cell Disease ◽

Sickle Cell ◽

Point Of Care ◽

Low Cost ◽

Testing Device ◽

Cell Disease ◽

Point Of Care Testing

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Label-Free Sickle Cell Disease Diagnosis using a Low-Cost, Handheld Platform

Advanced Materials Technologies ◽

10.1002/admt.201600100 ◽

2016 ◽

Vol 1 (5) ◽

pp. 1600100 ◽

Cited By ~ 22

Author(s):

Bekir Yenilmez ◽

Stephanie Knowlton ◽

Chu Hsiang Yu ◽

Matthew M. Heeney ◽

Savas Tasoglu

Keyword(s):

Sickle Cell Disease ◽

Sickle Cell ◽

Low Cost ◽

Disease Diagnosis ◽

Label Free ◽

Cell Disease

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Preliminary Evaluation of a Point-of-Care Testing Device (SickleSCAN™) in Screening for Sickle Cell Disease

Hemoglobin ◽

10.1080/03630269.2017.1329151 ◽

2017 ◽

Vol 41 (2) ◽

pp. 77-82 ◽

Cited By ~ 7

Author(s):

Maxwell M. Nwegbu ◽

Hezekiah A. Isa ◽

Biyaya B. Nwankwo ◽

Chinedu C. Okeke ◽

Uduak J. Edet-Offong ◽

...

Keyword(s):

Sickle Cell Disease ◽

Sickle Cell ◽

Point Of Care ◽

Preliminary Evaluation ◽

Testing Device ◽

Cell Disease ◽

Point Of Care Testing

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Point‐of‐Care Testing in Sickle Cell Disease

Sickle Cell Disease - Pain and Common Chronic Complications ◽

10.5772/64862 ◽

2016 ◽

Cited By ~ 2

Author(s):

Julie Kanter

Keyword(s):

Sickle Cell Disease ◽

Sickle Cell ◽

Point Of Care ◽

Cell Disease ◽

Point Of Care Testing

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Multicenter Evaluation of HemoTypeSC as a Point-of-Care Sickle Cell Disease Rapid Diagnostic Test for Newborns and Adults Across India

American Journal of Clinical Pathology ◽

10.1093/ajcp/aqz108 ◽

2019 ◽

Vol 153 (1) ◽

pp. 82-87 ◽

Cited By ~ 3

Author(s):

Malay B Mukherjee ◽

Roshan B Colah ◽

Pallavi R Mehta ◽

Nikhil Shinde ◽

Dipty Jain ◽

...

Keyword(s):

Sickle Cell Disease ◽

Newborn Screening ◽

Sickle Cell ◽

High Performance ◽

Screening Program ◽

Point Of Care ◽

Genetic Disorder ◽

Cell Disease ◽

Point Of Care Test ◽

Tribal Areas

Abstract Objectives Sickle cell anemia is the commonest genetic disorder in India, and the frequency of the sickle cell gene is very high in the remote tribal areas where facilities are generally limited. Therefore, a rapid and affordable point-of-care test for sickle cell disease is needed. Methods The diagnostic accuracy of HemoTypeSC was evaluated against automated high-performance liquid chromatography (HPLC) as the gold standard for its efficacy in a newborn screening program. Results A total of 1,559 individuals (980 newborns and 579 adults) from four participating centers were analyzed by both methods. HemoTypeSC correctly identified 209 of 211 total hemoglobin (Hb) SS cases, for a 99.1%/99.9% total HbSS sensitivity/specificity. Overall, HemoTypeSC exhibited sensitivity and specificity of 98.1% and 99.1% for all possible phenotypes (HbAA, HbAS, and HbSS) detected. HPLC is relatively expensive and not available in most laboratories in remote tribal areas. Conclusions We conclude that the rapid, point-of-care testing device HemoTypeSC test is suitable for population and newborn screening for the HbS phenotype.

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Newborn Screening for Sickle Cell Disease Using Point-of-Care Testing in Low-Income Setting

PEDIATRICS ◽

10.1542/peds.2018-4105 ◽

2019 ◽

Vol 144 (4) ◽

pp. e20184105 ◽

Cited By ~ 3

Author(s):

Ofelia A. Alvarez ◽

Tally Hustace ◽

Mimose Voltaire ◽

Alejandro Mantero ◽

Ulrick Liberus ◽

...

Keyword(s):

Sickle Cell Disease ◽

Newborn Screening ◽

Sickle Cell ◽

Low Income ◽

Point Of Care ◽

Cell Disease ◽

Point Of Care Testing ◽

Low Income Setting

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Erythrocytapheresis therapy to reduce iron overload in chronically transfused patients with sickle cell disease

Blood ◽

10.1182/blood.v83.4.1136.1136 ◽

1994 ◽

Vol 83 (4) ◽

pp. 1136-1142 ◽

Cited By ~ 95

Author(s):

HC Kim ◽

NP Dugan ◽

JH Silber ◽

MB Martin ◽

E Schwartz ◽

...

Keyword(s):

Sickle Cell Disease ◽

Iron Overload ◽

Sickle Cell ◽

Chelation Therapy ◽

Iron Chelation ◽

Cell Disease ◽

Donor Blood ◽

Iron Load ◽

Blood Usage ◽

Hb S

Abstract Chelation therapy with deferoxamine is effective in preventing the risk of transfusional iron overload, but treatment failure is common because of noncompliance. To reduce the transfusional iron load, we have evaluated longterm erythrocytapheresis in 14 subjects with sickle cell disease and stroke (11) or other complications (3) as an alternative to simple transfusion. Subjects were treated with erythrocytapheresis using the Haemonetics V50 (Haemonetics Corp, Braintree, MA) to maintain the target pretransfusion hemoglobin S (Hb S) level less than 50% for 6 to 71 months. The transfusional iron load and the donor blood usage were analyzed for a 6- to 36-month study period and were compared with similar data from a subset of 7 subjects previously treated with conventional (target Hb S < 30%) and modified (target Hb S < 50%) simple transfusion protocols. The effect of erythrocytapheresis on iron accumulation was determined by assessment of serum ferritin levels in the absence of iron chelation. The mean transfusional iron load and donor blood usage with erythrocytapheresis were 19 +/- 14 mg iron/kg/yr (range, 6 to 50) and 188.4 +/- 55.2 mL packed-red blood cells (RBC)/kg/yr (range, 107 to 281), respectively. Of 6 subjects receiving no iron chelation therapy, 5 maintained normal or nearly normal serum ferritin levels during 11 to 36 months of erythrocytapheresis. In comparison with conventional simple transfusion and modified simple transfusion, erythrocytapheresis reduced iron loading by 87% (P < .01) and 82% (P < .01), respectively, but increased donor blood usage by 23% and 73%, respectively. Subjects with pre-erythrocytapheresis Hb levels > or = 8.0 g/dL had lower iron accumulation (P < .001) and less donor blood usage (P < .005) than subjects with Hb levels < or = 8.0 g/dL. Although donor blood usage is increased in comparison with simple transfusion, long-term erythrocytapheresis markedly reduces or prevents iron accumulation. This form of transfusion therapy allows the cessation of iron chelation in well-chelated subjects and, if used as the initial form of transfusion therapy, may prevent long-term complications of sickle cell disease without risk of iron overload and the need for chelation therapy.

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Sickle cell disease resulting from uniparental disomy in a child who inherited sickle cell trait

Blood ◽

10.1182/blood-2010-05-284331 ◽

2010 ◽

Vol 116 (15) ◽

pp. 2822-2825 ◽

Cited By ~ 8

Author(s):

Jeffrey J. Swensen ◽

Archana M. Agarwal ◽

Jose M. Esquilin ◽

Sabina Swierczek ◽

Ajay Perumbeti ◽

...

Keyword(s):

Sickle Cell Disease ◽

Sickle Cell ◽

Sickle Cell Trait ◽

Uniparental Disomy ◽

Mendelian Inheritance ◽

Cell Disease ◽

Erythroid Progenitors ◽

Mendelian Genetics ◽

Abnormal Hemoglobin ◽

Hb S

Abstract Sickle cell disease (SCD) is a classic example of a disorder with recessive Mendelian inheritance, in which each parent contributes one mutant allele to an affected offspring. However, there are exceptions to that rule. We describe here the first reported case of conversion of inherited sickle cell trait to SCD by uniparental disomy (UPD) resulting in mosaicism for SS and AS erythrocytes. A 14-year-old boy presented with splenomegaly and hemolysis. Although his father has sickle cell trait, his mother has no abnormal hemoglobin (Hb). DNA sequencing, performed to rule out Hb S/β-thalassemia, detected homozygous Hb SS. Further studies revealed mosaic UPD of the β-globin locus, more SS erythroid progenitors than AS, but a reverse ratio of erythrocytes resulting from the survival advantage of AS erythrocytes. This report exemplifies non-Mendelian genetics wherein a patient who inherited sickle cell trait has mild SCD resulting from postzygotic mitotic recombination leading to UPD.

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Successful Outcome of Chronic Intrahepatic Cholestasis in an Adult Patient with Sickle Cell/β+Thalassemia

Case Reports in Hematology ◽

10.1155/2014/213631 ◽

2014 ◽

Vol 2014 ◽

pp. 1-3 ◽

Cited By ~ 1

Author(s):

Efthymia Vlachaki ◽

Panagiotis Andreadis ◽

Nikolaos Neokleous ◽

Aleka Agapidou ◽

Evaggelia Vetsiou ◽

...

Keyword(s):

Sickle Cell Disease ◽

Adult Patient ◽

Sickle Cell ◽

Intrahepatic Cholestasis ◽

Interesting Case ◽

Successful Outcome ◽

Variant Form ◽

Cell Disease ◽

Hb S ◽

Chronic Intrahepatic Cholestasis

Sickle cell/β+thalassemia (Hb S/β+thal) is considered as a variant form of sickle cell disease. Acute episodes of vasoocclusive pain crisis are characteristic for sickle cell disorders and may be complicated by an acute or chronic life-threatening organ dysfunction. Chronic intrahepatic cholestasis is a rare and severe complication in sickle cell disease, characterized by marked hyperbilirubinemia and acute hepatic failure with an often fatal course. Despite the fact that patients with Hb S/β+thal usually have a mild type of disease, herein we describe an interesting case of chronic intrahepatic cholestasis with successful outcome in an adult patient with Hb S/β+thal.

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