Detection of candidemia by polymerase chain reaction

Rand, Kenneth H.; Houck, Herbert; Wolff, Marcos

doi:10.1006/mcpr.1994.1030

Cited by 31 publications

(22 citation statements)

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“…Our results were in agreement with previous studies on the sensitivity of different PCR-based methodologies which indicate sensitivities of I-4 cfu/ml blood, 1 cfu/ml blood, 10 organisms/ 100 pl serum, 10 cfd200 pl serum and 10 organisms/l00 pl serum (22,24,25,27,28).…”

Section: Discussionsupporting

confidence: 93%

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Detection and identification of fungi in blood using broad‐range 28S rDNA PCR amplification and species‐specific hybridisation

Evertsson

Monstein

Johansson

2000

APMIS

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The aim of the present study was to develop a PCR-based method to detect and identify fungi directly from human venous blood. We used broad-range PCR primers that targeted a part of the large subunit 28S rRNA genes. To obtain species-specific hybridisation probes, type strains of Candida albicans, C. glabrata, C. krusei, C. parapsilosis, C. tropicalis and Cryptococcus neoformans were PCR amplified, and the amplicons were analysed by gene sequencing. Based on the sequence analysis, species-specific probes that targeted variable regions were designed and used in hybridisation analyses. Between 2 to 10 fungal cells/ml of spiked blood samples could be detected and correctly identified to species. We applied the technique to blood samples obtained from two patients with or two patients without verified candidaemia. The three samples of candidaemia patients were correctly identified to species level, and those of the negative patients remained negative. This method is a potential tool for diagnosis of systemic invasive candidiasis.

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Section: Discussionsupporting

confidence: 93%

“…Some of these methods, however, detect only C. albicans (10,22,23) or just a few of the non-C. albicans known to cause candidaemia (22). Certain methods are based on PCR-amplification of low-copy genes (24,25), which probably does not favour sensitivity.…”

Section: Discussionmentioning

confidence: 99%

Detection and identification of fungi in blood using broad‐range 28S rDNA PCR amplification and species‐specific hybridisation

Evertsson

Monstein

Johansson

2000

APMIS

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“…C. albicans was detected in seeded blood specimens (18,81,128), in blood samples from experimentally infected animals (95,200), and in human blood in one study (95). The sensitivity of the PCR for C. albicans was disappointing: 79% (95), 73% (26), and 46% (158). Two possible reasons for this lack of sensitivity have been mentioned: the difficulty in releasing DNA from C. albicans cells, a critical need when they are present in small numbers (162); and the small volume of the specimen used in the amplification reaction (158).…”

Section: Fungimentioning

confidence: 99%

“…The sensitivity of the PCR for C. albicans was disappointing: 79% (95), 73% (26), and 46% (158). Two possible reasons for this lack of sensitivity have been mentioned: the difficulty in releasing DNA from C. albicans cells, a critical need when they are present in small numbers (162); and the small volume of the specimen used in the amplification reaction (158). PCR has been used more frequently for classification and identification of Candida spp.…”

Section: Fungimentioning

confidence: 99%

Relevance of nucleic acid amplification techniques for the diagnosis of respiratory tract infections in the clinical laboratory

Ieven¹

1996

Journal of Microbiological Methods

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“…Conventional blood cultures are not very effective in detecting yeast growth and take too long to become positive (26), although great progress has been made in recent years (7). Alternative procedures such as the detection of circulating antigens, enzymes, or metabolic products (17,26,31,33) lack sensitivity and, to some extent, specificity, so diagnosis can be delayed or in most cases is obtained only at autopsy (4,8,10,25,26). Since candidosis is the most common disseminated fungal infection in leukemia or cancer patients, an early diagnosis of candidemia is therefore of great importance for patient care, especially if it leads to the selection of a rapid, species-specific antifungal therapy, which could likely reduce mortality (8).…”

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confidence: 99%

Identification of various medically important Candida species in clinical specimens by PCR-restriction enzyme analysis

et al. 1997

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A single primer pair amplifying a cytochrome P-450 lanosterol-14␣-demethylase (L1A1) gene fragment that encodes a highly conserved region was used to detect yeast DNA in clinical specimens. Positive PCR products were obtained from genomic DNAs of Candida albicans, C. parapsilosis, C. tropicalis, C. guilliermondii, C. krusei, C. (Torulopsis) glabrata, and C. kefyr. No human, bacterial, or parasitic DNA was amplified. The sensitivity was evaluated for C. albicans genomic DNA by using various DNA concentrations (200 pg to 2 fg). The amplified DNAs of Candida species with unknown P-450 L1A1 gene sequences were subcloned and sequenced. Identification at the species level was achieved by digestion of the PCR products with different restriction enzymes. A specific restriction enzyme analysis pattern was determined for each species investigated. Subsequently, we used PCR to detect specific yeast DNA directly with clinical specimens such as blood and bronchoalveolar lavage specimens. After appropriate treatment, the specimens were processed by PCR and the results were compared with those obtained by traditional diagnostic procedures such as cultures and serology. Although preliminary, the PCR results seem to correlate well, at least for blood, with those of antigen detection assays and traditional blood cultures, with a better and earlier detection of candidemia.

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Detection of candidemia by polymerase chain reaction

Cited by 31 publications

References 0 publications

Detection and identification of fungi in blood using broad‐range 28S rDNA PCR amplification and species‐specific hybridisation

Detection and identification of fungi in blood using broad‐range 28S rDNA PCR amplification and species‐specific hybridisation

Relevance of nucleic acid amplification techniques for the diagnosis of respiratory tract infections in the clinical laboratory

Identification of various medically important Candida species in clinical specimens by PCR-restriction enzyme analysis

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