Evaluation of the LightCycler® PRO Instrument as a Platform for Rhesus D Typing

Abstract

Rapid and reliable Rhesus D typing is crucial for blood donation centers. In instances of massive blood transfusion or reduced antigen expression, DNA-based phenotype prediction becomes mandatory. Our molecular RHD typing approach involves an initial real-time PCR for the most common aberrant RHD types in our region, RHD*01W.1 (weak D type 1), RHD*01W.2 (weak D type 2), RHD*01W.3 (weak D type 3), and RHD*07.01 (DVII). For comprehensive coverage, Sanger sequencing of RHD coding regions is performed in the case of PCR target-negative results. We evaluated the specificity and accuracy of these methods using the recently launched LightCycler^® PRO real-time platform. All findings demonstrated remarkable accuracy. Notably, the LightCycler^® PRO instrument offers a distinct advantage in data interpretation and integration via the HL7 interface. This study underlines the importance of including advanced molecular techniques in blood typing protocols, especially in scenarios where conventional serological methods may be insufficient.

Keywords: LightCycler® PRO; RHD typing; Rhesus D antigen; blood group genotyping; transfusion medicine.

Figure 1

Specific melting curves of (a) RHD*01W.1, (b) RHD*01W.2, (c) RHD*01W.3, and (d) RHD*07.01 PCR reactions. Different color lines represent distinct samples.

Figure 2

Melting Curve Genotyping analysis report of RHD*01W.1-specific PCR reaction.