ArticleClinical and diagnostic laboratory immunology2002
Detection of Cryptococcus neoformans DNA in tissue samples by nested and real-time PCR assays.
Article in Clinical and diagnostic laboratory immunology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 63 citations in OpenAlex.
- Evolution of Laboratory Diagnostics for Cryptococcosis and Missing Links to Optimize Diagnosis and Outcomes in Resource-Constrained Settings.Open forum infectious diseases · 2024Review
- Comparative analysis of diagnostic methods for the detection of Cryptococcus neoformans meningitis.PLoS neglected tropical diseases · 2023Article
- Diagnostic and therapeutic management of Cryptococcosis in a kitten with practical considerations to veterinary pediatric therapeutic approach.Medical mycology case reports · 2021Article
- Identification of Fungal Species and Detection of Azole-Resistance Mutations in theYonsei medical journal · 2020Article
- Comparison of Antigen Detection and Nested PCR in CSF Samples of HIV Positive and Negative Patients with Suspected Cryptococcal Meningitis in a Tertiary Care Hospital.Journal of clinical and diagnostic research : JCDR · 2016Article
- Cryptococcus gattii infections.Clinical microbiology reviews · 2014Review
- Importance of the association of molecular and immunological diagnosis in immunocompetent patient with Histoplasma capsulatum and Cryptoccocus neoformans infection: a case report.The journal of venomous animals and toxins including tropical diseases · 2014Article
- A multi-target real-time PCR assay for rapid identification of meningitis-associated microorganisms.Molecular biotechnology · 2013Article
- Identification of fungal species by fragment length analysis of the internally transcribed spacer 2 region.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2009Article
- Granulomatous prostatitis due to Cryptococcus neoformans: diagnostic usefulness of special stains and molecular analysis of 18S rDNA.Prostate cancer and prostatic diseases · 2008Article
- Real-time PCR in clinical microbiology: applications for routine laboratory testing.Clinical microbiology reviews · 2006Review
- Diagnosis of invasive aspergillosis and mucormycosis in immunocompromised patients by seminested PCR assay of tissue samples.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2006Article
- Rapid identification and enumeration of Saccharomyces cerevisiae cells in wine by real-time PCR.Applied and environmental microbiology · 2005Article
- PCR based identification and discrimination of agents of mucormycosis and aspergillosis in paraffin wax embedded tissue.Journal of clinical pathology · 2005Article
- Use of a suspension array for rapid identification of the varieties and genotypes of the Cryptococcus neoformans species complex.Journal of clinical microbiology · 2005Article
- PCR assays for identification of Coccidioides posadasii based on the nucleotide sequence of the antigen 2/proline-rich antigen.Journal of clinical microbiology · 2004Article
- Sensitive and quantitative detection of severe acute respiratory syndrome coronavirus infection by real-time nested polymerase chain reaction.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2004Article
- Application of real-time polymerase chain reaction technology to detect prostatic bacteria in patients with chronic prostatitis/chronic pelvic pain syndrome.World journal of urology · 2003Article
- Detection of single cells of Cryptococcus neoformans in clinical samples by solid-phase cytometry.Journal of clinical microbiology · 2003Article
- Nested PCR assays for detection of Blastomyces dermatitidis DNA in paraffin-embedded canine tissue.Journal of clinical microbiology · 2003Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Two PCR protocols targeting the 18S rRNA gene of Cryptococcus neoformans were established, compared, and evaluated in murine cryptococcal meningitis. One protocol was designed as a nested PCR to be performed in conventional block thermal cyclers. The other protocol was designed as a quantitative single-round PCR adapted to LightCycler technology. One hundred brain homogenates and dilutions originating from 20 ICR mice treated with different azoles were examined. A fungal burden of 3 x 10(1) to 2.9 x 10(4) CFU per mg of brain tissue was determined by quantitative culture. Specific PCR products were amplified by the conventional and the LightCycler methods in 86 and 87 samples, respectively, with products identified by DNA sequencing and real-time fluorescence detection. An analytical sensitivity of 1 CFU of C. neoformans per mg of brain tissue and less than 10 CFU per volume used for extraction was observed for both PCR protocols, while homogenates of 70 organs from mice infected with other fungi were PCR negative. Specificity testing was performed with genomic DNA from 31 hymenomycetous fungal species and from the ustilaginomycetous yeast Malassezia furfur, which are phylogenetically related to C. neoformans. Twenty-four strains, including species of human skin flora like M. furfur and Trichosporon spp., were PCR negative. Amplification was observed with Cryptococcus amylolentus, Filobasidiella depauperata, Cryptococcus laurentii, and five species unrelated to clinical specimens. LightCycler PCR products from F. depauperata and Trichosporon faecale could be clearly discriminated by melting curve analysis. The sensitive and specific nested PCR assay as well as the rapid and quantitative LightCycler PCR assay might be useful for the diagnosis and monitoring of human cryptococcal infections.
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