Single nucleotide polymorphism profiling assay to exclude serum sample mix-up

Vox Sang. 2007 Feb;92(2):148-53. doi: 10.1111/j.1423-0410.2006.00871.x.

Abstract

Background and objectives: Sample mix-ups are a threat to the validity of clinical laboratory test results. To detect serum sample mix-ups we developed a single nucleotide polymorphism (SNP) profiling test. SNPs are frequent sequence variations in the human genome. Each individual has a unique combination of these nucleotide variations.

Materials and methods: Predeveloped SNP amplification assays are commercially available. We recently discovered that these SNP assays could be applied to serological samples, which is not self-evident because a key step in serum preparation is removal of white blood cells, the major source of DNA, from blood. DNA was extracted from serum samples. Real-time polymerase chain reaction (PCR) analysis of the purified DNA using a selection of 10 SNP assays provided SNP profiles.

Results: The applicability of the SNP profiling test was demonstrated by means of a case where hepatitis E virus serological determinations of four serum samples of one patient seemed inconsistent. SNP profiling of the samples demonstrated that this was due to the enzyme-linked immunosorbent assay test instead of sample mix-up.

Conclusion: We have developed an SNP profiling assay that provides a way to link human serum samples to a source, without post-PCR processing. The chance for two randomly chosen individuals to have an identical profile is 1 in 18 000. Solving potential serum sample mix-ups will secure downstream evaluations and critical decisions concerning the patients involved.

MeSH terms

  • Clinical Laboratory Techniques / methods*
  • DNA / analysis*
  • Diagnostic Errors / prevention & control*
  • Gene Frequency
  • Hepatitis E / blood
  • Hepatitis E virus / genetics
  • Hepatitis E virus / immunology
  • Hepatitis E virus / isolation & purification
  • Humans
  • Polymorphism, Single Nucleotide*
  • Quality Control*
  • Serum / chemistry
  • Specimen Handling

Substances

  • DNA