Development of DNA probes for Candida albicans

Cheung, Lori L.; Hudson, Jane

doi:10.1016/0732-8893(88)90037-5

Cited by 19 publications

(16 citation statements)

References 7 publications

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“…This is also illustrated by the fact that the majority of published case reports have been severely criticised as being poorly substantiated [28,29]. Recently, however, we have substantiated two cases of cryptococcaemia and cryptococcal meningitis in AIDS patients, which were due to these two cryptococcal species 1 [30]. The method described here can, therefore, be of potential use in the prompt identi¢cation of C. albidus and C. laurentii, as serum LPA tests are negative in patients infected by these two organisms and traditional methods of identi¢cation are time consuming [30].…”

Section: Discussionmentioning

confidence: 82%

Identification of medically significant fungal genera by polymerase chain reaction followed by restriction enzyme analysis

Velegraki¹

1999

FEMS Immunology and Medical Microbiology

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Rapid non-culture-dependent assays for identification of fungi quicken diagnosis and prompt treatment of invasive fungal disease. Fungal DNA extracts from pure cultures of the most frequently isolated fungal pathogens belonging to the Genera Aspergillus, Candida and Cryptococcus along with less common pathogenic Genera were amplified with the general fungal primer pair internal transcribed spacer-1/4. Subsequently, the amplicon was digested with the restriction endonucleases MspI, HaeIII, HinfI and EcoRI in order to generate genus- or species-specific patterns for identification of the fungus. HinfI produced indistinguishable fingerprints for all Aspergillus species tested. MspI produced species-specific patterns for: Cryptococcus neoformans, Cryptococcus non-neoformans, Candida albicans and Candida tropicalis. EcoRI succeeded in differentiating penicillia from aspergilli and cryptococci from Candida spp. It is concluded that this procedure can differentiate genera and occasionally species of medically important fungi and that following the necessary validation experiments, it can be used directly on clinical samples to assist prompt diagnosis of systemic fungal infections.

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Section: Discussionmentioning

confidence: 82%

Identification of medically significant fungal genera by polymerase chain reaction followed by restriction enzyme analysis

Velegraki¹

1999

FEMS Immunology and Medical Microbiology

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“…(145,154). Although the entire procedure is rapid and easy to perform, the sensitivity of the assay is often lower than those of other molecular biological assays, especially those including nucleic acid amplification (39,87,171).…”

Section: Amplification and Detection Methodsmentioning

confidence: 99%

Current Status of Nonculture Methods for Diagnosis of Invasive Fungal Infections

2002

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The incidence of invasive fungal infections has increased dramatically in recent decades, especially among immunocompromised patients. However, the diagnosis of these infections in a timely fashion is often very difficult. Conventional microbiologic and histopathologic approaches generally are neither sensitive nor specific, and they often do not detect invasive fungal infection until late in the course of disease. Since early diagnosis may guide appropriate treatment and prevent mortality, there has been considerable interest in developing nonculture approaches to diagnosing fungal infections. These approaches include detection of specific host immune responses to fungal antigens, detection of specific macromolecular antigens using immunologic reagents, amplification and detection of specific fungal nucleic acid sequences, and detection and quantitation of specific fungal metabolite products. This work reviews the current status and recent developments as well as problems in the design of nonculture diagnostic methods for invasive fungal infections

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“…In order to overcome the limitations of conventional diagnostic tests, DNA-based methods have been developed for the detection of Candida species and offer a potentially more sensitive means of diagnosing systemic candidiasis (4,30). The use of PCR-based tests to detect Candida DNA in body fluids has produced encouraging results (5,13,15,16,23).…”

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confidence: 99%

Seminested PCR for Diagnosis of Candidemia: Comparison with Culture, Antigen Detection, and Biochemical Methods for Species Identification

et al. 2002

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The rapid detection and identification of Candida species in clinical laboratories are extremely important for the management of patients with hematogenous candidiasis. The presently available culture and biochemical methods for detection and species identification of Candida are time-consuming and lack the required sensitivity and specificity. In this study, we have established a seminested PCR (snPCR) using universal and species-specific primers for detection of Candida species in serum specimens. The universal outer primers amplified the 3 end of 5.8S ribosomal DNA (rDNA) and the 5 end of 28S rDNA, including the internally transcribed spacer 2 (ITS2), generating 350-to 410-bp fragments from the four commonly encountered Candida species, viz., C. albicans, C. tropicalis, C. glabrata, and C. parapsilosis. The species-specific primers, complementary to unique sequences within the ITS2 of each test species, amplified species-specific DNA in the reamplification step of the snPCR. The sensitivity of Candida detection by snPCR in spiked serum specimens was close to 1 organism/ml. Evaluation of snPCR for specific identification of Candida species with 76 clinical Candida isolates showed 99% concordant results with the Vitek and/or ID32C yeast identification system. Further evaluation of snPCR for detection of Candida species in sera from culture-proven (n ‫؍‬ 12), suspected (n ‫؍‬ 16), and superficially colonized (n ‫؍‬ 10) patients and healthy subjects (n ‫؍‬ 12) showed that snPCR results were consistently negative with sera from healthy individuals and colonized patients. In culture-proven candidemia patients, the snPCR results were in full agreement with blood culture results with respect to both positivity and species identity. In addition, snPCR detected candidemia due to two Candida species in five patients, compared to three by blood culture. In the category of suspected candidemia with negative blood cultures for Candida, nine patients (56%) were positive by snPCR; two of them had dual infection with C. albicans and either C. tropicalis or C. glabrata. In conclusion, the snPCR developed in this study is specific and more sensitive than culture for the detection of Candida species in serum specimens. Moreover, the improved detection of cases of candidemia caused by more than one Candida species is an additional advantage.

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Development of DNA probes for Candida albicans

Cited by 19 publications

References 7 publications

Identification of medically significant fungal genera by polymerase chain reaction followed by restriction enzyme analysis

Identification of medically significant fungal genera by polymerase chain reaction followed by restriction enzyme analysis

Current Status of Nonculture Methods for Diagnosis of Invasive Fungal Infections

Seminested PCR for Diagnosis of Candidemia: Comparison with Culture, Antigen Detection, and Biochemical Methods for Species Identification

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