Application of an oligonucleotide microarray-based nano-amplification technique for the detection of fungal pathogens
-
Weiping Lu
Abstract
Background: The traditional techniques for diagnosis of invasive fungal infections in the clinical microbiology laboratory need improvement. These techniques are prone to delay results due to their time-consuming process, or result in misidentification of the fungus due to low sensitivity or low specificity. The aim of this study was to develop a method for the rapid detection and identification of fungal pathogens.
Methods: The internal transcribed spacer two fragments of fungal ribosomal DNA were amplified using a polymerase chain reaction for all samples. Next, the products were hybridized with the probes immobilized on the surface of a microarray. These species-specific probes were designed to detect nine different clinical pathogenic fungi including Candida albicans, Candida tropocalis, Candida glabrata, Candida parapsilosis, Candida krusei, Candida lusitaniae, Candida guilliermondii, Candida keyfr, and Cryptococcus neoformans. The hybridizing signals were enhanced with gold nanoparticles and silver deposition, and detected using a flatbed scanner or visually.
Results: Fifty-nine strains of fungal pathogens, including standard and clinically isolated strains, were correctly identified by this method. The sensitivity of the assay for Candida albicans was 10 cells/mL. Ten cultures from clinical specimens and 12 clinical samples spiked with fungi were also identified correctly.
Conclusions: This technique offers a reliable alternative to conventional methods for the detection and identification of fungal pathogens. It has higher efficiency, specificity and sensitivity compared with other methods commonly used in the clinical laboratory.
Clin Chem Lab Med 2010;48:1507–14.
©2010 by Walter de Gruyter Berlin New York
Articles in the same Issue
- Editorial
- Darwinian evolution or regression? The fate of laboratory professionals
- Review
- Automated reticulocyte counting: state of the art and clinical applications in the evaluation of erythropoiesis
- Minireview
- Laboratory diagnostics in acute poisoning: critical overview
- Opinion Papers
- On the origins of physicians: Darwinian or Lamarckian evolution?
- Observed, unknown distributions of clinical chemical quantities should be considered to be log-normal: a proposal
- Guidelines and Recommendations
- Development and description of GETT: a Genetic testing Evidence Tracking Tool
- Genetics and Molecular Diagnostics
- DNA sequencing errors in molecular diagnostics of filamin myopathy
- High resolution melting for the identification of mutations in the iron responsive element of the ferritin light chain gene
- General Clinical Chemistry and Laboratory Medicine
- Detection of serum free light chains: the problem with antigen excess
- Hepcidin concentrations and iron homeostasis in preeclampsia
- Automated assay for non-transferrin-bound iron in serum samples
- A simple liquid chromatography-tandem mass spectrometry method for urinary free cortisol analysis: suitable for routine purpose
- Evaluation of a multiplex assay for adipokine concentrations in obese children
- The use of serial patient blood gas, electrolyte and glucose results to derive biologic variation: a new tool to assess the acceptability of intensive care unit testing
- Point-of-care determination of neonatal bilirubin with the blood gas analyzer RapidLab 1265
- The effect of pre-analytical variables on light transmittance aggregometry in citrated platelet-rich plasma from healthy subjects
- Cancer Diagnostics
- Clinical significance of serum macrophage-colony stimulating factor (M-CSF) in esophageal cancer patients and its comparison with classical tumor markers
- VKORC1 haplotypes influence the performance characteristics of PIVKAII for screening of hepatocellular carcinoma
- Serum soluble tumour necrosis factor receptor type I concentrations independently predict prognosis in patients with breast cancer
- Variations in systemic biomarkers of oxidative/nitrosative stress and DNA damage before and during the consequent two cycles of chemotherapy in breast cancer patients
- Spurious increase in serum chromogranin A: the role of heterophilic antibodies
- Infectious Diseases
- Identification and evaluation of a new nucleic acid amplification test target for specific detection of Mycobacterium tuberculosis
- Application of an oligonucleotide microarray-based nano-amplification technique for the detection of fungal pathogens
- Liquid chromatography-tandem mass spectrometry method for simultaneous quantification of four triazole antifungal agents in human plasma
- Evaluation of two sirolimus assays using the ARCHITECT-i1000® CMIA or RxL® ACMIA methods in comparison with the IMx® MEIA method
- Widespread use of point-of-care testing is irreconcilable with the present-day quest for safety
- Congress Abstracts
- 3rd Annual Congress of the Austrian Society for Laboratory Medicine and Clinical Chemistry Salzburg, Austria, 27-30 October 2010
Articles in the same Issue
- Editorial
- Darwinian evolution or regression? The fate of laboratory professionals
- Review
- Automated reticulocyte counting: state of the art and clinical applications in the evaluation of erythropoiesis
- Minireview
- Laboratory diagnostics in acute poisoning: critical overview
- Opinion Papers
- On the origins of physicians: Darwinian or Lamarckian evolution?
- Observed, unknown distributions of clinical chemical quantities should be considered to be log-normal: a proposal
- Guidelines and Recommendations
- Development and description of GETT: a Genetic testing Evidence Tracking Tool
- Genetics and Molecular Diagnostics
- DNA sequencing errors in molecular diagnostics of filamin myopathy
- High resolution melting for the identification of mutations in the iron responsive element of the ferritin light chain gene
- General Clinical Chemistry and Laboratory Medicine
- Detection of serum free light chains: the problem with antigen excess
- Hepcidin concentrations and iron homeostasis in preeclampsia
- Automated assay for non-transferrin-bound iron in serum samples
- A simple liquid chromatography-tandem mass spectrometry method for urinary free cortisol analysis: suitable for routine purpose
- Evaluation of a multiplex assay for adipokine concentrations in obese children
- The use of serial patient blood gas, electrolyte and glucose results to derive biologic variation: a new tool to assess the acceptability of intensive care unit testing
- Point-of-care determination of neonatal bilirubin with the blood gas analyzer RapidLab 1265
- The effect of pre-analytical variables on light transmittance aggregometry in citrated platelet-rich plasma from healthy subjects
- Cancer Diagnostics
- Clinical significance of serum macrophage-colony stimulating factor (M-CSF) in esophageal cancer patients and its comparison with classical tumor markers
- VKORC1 haplotypes influence the performance characteristics of PIVKAII for screening of hepatocellular carcinoma
- Serum soluble tumour necrosis factor receptor type I concentrations independently predict prognosis in patients with breast cancer
- Variations in systemic biomarkers of oxidative/nitrosative stress and DNA damage before and during the consequent two cycles of chemotherapy in breast cancer patients
- Spurious increase in serum chromogranin A: the role of heterophilic antibodies
- Infectious Diseases
- Identification and evaluation of a new nucleic acid amplification test target for specific detection of Mycobacterium tuberculosis
- Application of an oligonucleotide microarray-based nano-amplification technique for the detection of fungal pathogens
- Liquid chromatography-tandem mass spectrometry method for simultaneous quantification of four triazole antifungal agents in human plasma
- Evaluation of two sirolimus assays using the ARCHITECT-i1000® CMIA or RxL® ACMIA methods in comparison with the IMx® MEIA method
- Widespread use of point-of-care testing is irreconcilable with the present-day quest for safety
- Congress Abstracts
- 3rd Annual Congress of the Austrian Society for Laboratory Medicine and Clinical Chemistry Salzburg, Austria, 27-30 October 2010
