Molecular Biology Reports

, Volume 46, Issue 2, pp 2379–2385 | Cite as

Real-time PCR based detection of the lactase non-persistence associated genetic variant LCT-13910C>T directly from whole blood

  • Axel MuendleinEmail author
  • Andreas Leiherer
  • Christina Zach
  • Eva Maria Brandtner
  • Peter Fraunberger
  • Heinz Drexel
  • Kathrin Geiger
Original Article


Primary hypolactasia is the main cause of lactose intolerance in adults. It is strongly associated with the single genetic variant LCT-13910C>T, located upstream of the lactase encoding gene. Consequently, analysis of LCT-13910C>T has been recommended as a direct genetic test for the trait. The aim of our study was to develop a TaqMan probe based real-time PCR protocol for the detection of the LCT-13910C>T variant directly from whole blood, circumventing DNA isolation. The LCT-13910C>T variant was determined using the DirectBlood Genotyping PCR Kit (myPOLS Biotec, Konstanz, Germany) together with an in-house TaqMan primer-probe assay. Validity and specificity of the assay was evaluated using EDTA anti-coagulated whole blood samples and corresponding DNA samples. Results from real-time PCR were compared with results obtained by Sanger sequencing from 105 blinded whole blood samples. Validity and specificity of the assay using whole blood were comparable to those using purified genomic DNA as substrate in PCR. Genetic analysis of blood samples were in complete agreement with results obtained by Sanger sequencing. In conclusion, we present a reliable real-time PCR protocol for the detection of the LCT-13910C>T variant directly from whole blood further facilitating diagnosis of primary hypolactasia in symptomatic patients.


Lactose intolerance Primary hypolactasia LCT-13910C>T Real-time PCR Direct blood PCR TaqMan probe Locked nucleic acids 



We are grateful to the Vorarlberger Landesregierung (Bregenz, Austria) for continuously supporting our research institute.


This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest related to the contents of this article.

Informed consent

All patients gave written confirmed consent for clinical LCT-13910C>T testing.

Supplementary material

11033_2019_4696_MOESM1_ESM.xlsx (14 kb)
Supplementary material 1 (XLSX 14 KB)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Axel Muendlein
    • 1
    Email author
  • Andreas Leiherer
    • 1
    • 2
    • 3
  • Christina Zach
    • 1
    • 2
  • Eva Maria Brandtner
    • 1
  • Peter Fraunberger
    • 2
  • Heinz Drexel
    • 1
    • 3
    • 4
    • 5
  • Kathrin Geiger
    • 1
  1. 1.Vorarlberg Institute for Vascular Investigation and Treatment (VIVIT)FeldkirchAustria
  2. 2.Medical Central LaboratoriesFeldkirchAustria
  3. 3.Private University of the Principality of LiechtensteinTriesenLiechtenstein
  4. 4.Division of Angiology, Swiss Cardiovascular CenterUniversity Hospital of BerneBerneSwitzerland
  5. 5.Drexel University College of MedicinePhiladelphiaUSA

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