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Innovative PCR without DNA extraction for African sickle cell disease diagnosis.

L Detemmerman1, S Olivier1, V Bours2

  • 1a LaCAR MDX Technologies , Liège , Belgium.

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|September 21, 2017
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Summary

A new molecular diagnostic test accurately detects hemoglobin S (HbS) and hemoglobin C (HbC) directly from blood samples, including dried blood spots. This method simplifies diagnosis for Hb disorders, reducing costs and turnaround time.

Keywords:
African countriesHbCHbSMolecular diagnosisPCRdried blood spotnewborn screeningsickle cell disease

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Area of Science:

  • Molecular Diagnostics
  • Hematology
  • Genetics

Background:

  • Hemoglobin (Hb) disorders, including thalassemia and structural variants like HbS and HbC, cause severe anemia and vascular issues.
  • Diagnosis typically relies on advanced lab techniques (isoelectric focusing, HPLC, mass spectrometry), which are not widely accessible, especially in sub-Saharan Africa.
  • Accurate and accessible Hb disorder diagnosis is crucial for managing associated health complications.

Purpose of the Study:

  • To evaluate the performance of a novel molecular diagnostic test, the Human Hb S/C Lamp assay, for detecting HbS and HbC alleles.
  • To assess the feasibility of using this assay on various sample types, including dried blood spots (DBS), without prior DNA extraction.
  • To compare the LAMP assay's results against established diagnostic methods.

Main Methods:

  • The Human Hb S/C Lamp assay, a polymerase chain reaction-based method, was used to genotype 248 blood samples (whole blood and DBS).
  • The assay detects HbS, HbC, and HbA alleles directly from samples, eliminating the need for DNA extraction.
  • Genotyping results from the LAMP assay were compared with routine diagnostic methods at a university hospital genetics lab.

Main Results:

  • The LAMP assay demonstrated 100% accuracy in detecting both HbS and HbC alleles.
  • No interference was observed with fetal hemoglobin, indicating suitability for neonatal screening.
  • The assay successfully genotyped samples directly from fresh/frozen blood and DBS.

Conclusions:

  • The Human Hb S/C Lamp assay offers a highly accurate and accessible molecular diagnostic method for HbS and HbC.
  • This assay is unique in its ability to perform molecular testing directly on dried blood cards without DNA extraction.
  • The method significantly reduces handling time, turnaround time, and costs associated with diagnosing Hb disorders.