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Harmonization of PCR-based detection of intestinal pathogens: experiences from the Dutch external quality assessment scheme on molecular diagnosis of protozoa in stool samples
- Source :
- Clinical Chemistry and Laboratory Medicine (CCLM), 56(10), 1722-1727, Clinical Chemistry and Laboratory Medicine, 56, 1722-1727, Clinical Chemistry and Laboratory Medicine, 56, 10, pp. 1722-1727, Clinical Chemistry and Laboratory Medicine, 56(10), 1722-1727. De Gruyter
- Publication Year :
- 2018
-
Abstract
- Background: Real-time PCR methods are increasingly used in routine patient care settings not only to determine the presence or absence of pathogens in patient materials, but also to obtain semiquantitative results to estimate the pathogen load. However, it is so far unknown how well these methods are harmonized among different laboratories. Methods: Sets of stool samples were distributed three to four times per year to ca. 25–40 participating laboratories within the European Union as part of an external quality assessment scheme (EQAS) for the detection of gastrointestinal protozoa. This paper presents the results obtained over a 3-year period for Entamoeba histolytica, Entamoeba dispar, Giardia lamblia, Cryptosporidium species and Dientamoeba fragilis. Results: Although both false-positive and false-negative results were reported, the overall sensitivity and specificity were high. The substantial differences in the quantitative output of the real-time PCR assays could be traced back to differences in DNA isolation procedures between different laboratories. Conclusions: Participation in an EQAS proved to be important as it provides information on how the real-time PCR methods used by the participant compares to the generally reported results and indicates how procedures could be improved. Semiquantitative results of real-time PCR methods are not exchangeable between laboratories as long as the diagnostic procedures are not harmonized. Intralaboratory comparison of semiquantitative real-time PCR results seems only possible by the use of calibration curves derived from well-validated standards in clinical material and not by spiking solutions with purified DNA.
- Subjects :
- Quality Control
0301 basic medicine
medicine.medical_specialty
030106 microbiology
030231 tropical medicine
Clinical Biochemistry
Dientamoeba fragilis
Cryptosporidium
external quality assessment scheme
Biology
medicine.disease_cause
Polymerase Chain Reaction
Sensitivity and Specificity
Feces
03 medical and health sciences
Entamoeba histolytica
0302 clinical medicine
All institutes and research themes of the Radboud University Medical Center
Internal medicine
External quality assessment
Parasitic Diseases
medicine
Animals
Humans
Giardia lamblia
media_common.cataloged_instance
Parasites
European union
Netherlands
media_common
intestinal protozoa
Intralaboratory
Biochemistry (medical)
General Medicine
DNA, Protozoan
Laboratories, Hospital
biology.organism_classification
DNA extraction
Gastrointestinal Tract
lnfectious Diseases and Global Health Radboud Institute for Health Sciences [Radboudumc 4]
real-time PCR
Subjects
Details
- ISSN :
- 14346621
- Database :
- OpenAIRE
- Journal :
- Clinical Chemistry and Laboratory Medicine (CCLM), 56(10), 1722-1727, Clinical Chemistry and Laboratory Medicine, 56, 1722-1727, Clinical Chemistry and Laboratory Medicine, 56, 10, pp. 1722-1727, Clinical Chemistry and Laboratory Medicine, 56(10), 1722-1727. De Gruyter
- Accession number :
- edsair.doi.dedup.....9f58ad7ae4f0e309df9faa8beb7c104d