PCR-enzyme immunoassay of rDNA in the diagnosis of candidemia and comparison with amplicon detection by agarose gel electrophoresis

Ahmad, Suhail; Mustafa, Abu Salim; Khan, ZU; Al-Rifaiy, Areej I; Khan, Zia U.

doi:10.1016/j.ijmm.2004.01.002

Cited by 43 publications

(44 citation statements)

References 23 publications

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“…Multiple studies have evaluated the performance of PCR tests for the diagnosis of invasive Candida infections in patient populations (4,118,(185)(186)(187)(188)(189)(190)(191)(192)(193)(194)(195)(196)(197)(198)(199)(200)(201)(202) (studies are summarized in Table 5). The ranges of reported sensitivity and specificity values are 56.2 to 100% and 54 to 100%, respectively.…”

Section: Pcrmentioning

confidence: 99%

“…These differences should be considered fundamental, as accuracy values may change significantly depending on the way that each study handles cases of probable and possible invasive candidiasis. Interestingly, many studies show a tendency of PCR methods to detect Candida DNA in patients at high risk of invasive candidiasis with negative blood culture (187,189,202). In some cases of discordant results between blood cultures and PCR, Candida spp.…”

Section: Pcrmentioning

confidence: 99%

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Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

et al. 2014

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SUMMARY Invasive fungal infections constitute a serious threat to an ever-growing population of immunocompromised individuals and other individuals at risk. Traditional diagnostic methods, such as histopathology and culture, which are still considered the gold standards, have low sensitivity, which underscores the need for the development of new means of detecting fungal infectious agents. Indeed, novel serologic and molecular techniques have been developed and are currently under clinical evaluation. Tests like the galactomannan antigen test for aspergillosis and the β-glucan test for invasive Candida spp. and molds, as well as other antigen and antibody tests, for Cryptococcus spp., Pneumocystis spp., and dimorphic fungi, have already been established as important diagnostic approaches and are implemented in routine clinical practice. On the other hand, PCR and other molecular approaches, such as matrix-assisted laser desorption ionization (MALDI) and fluorescence in situ hybridization (FISH), have proved promising in clinical trials but still need to undergo standardization before their clinical use can become widespread. The purpose of this review is to highlight the different diagnostic approaches that are currently utilized or under development for invasive fungal infections and to identify their performance characteristics and the challenges associated with their use.

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Section: Pcrmentioning

confidence: 99%

Section: Pcrmentioning

confidence: 99%

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

et al. 2014

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“…In order to overcome these difficulties, non-culture methods have been developed. With these methods, highly sensitive and specific detection of fungal DNA from clinical materials has been demonstrated (Maaroufi et al, 2003;Ahmad et al, 2004). However, technical problems such as the risk of contamination and the inability to distinguish colonized cases from infected patients may need to be solved.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of a newly developed down-flow immunoassay for detection of serum mannan antigens in patients with candidaemia

Fujita¹,

Takamura²,

Nagahara³

et al. 2006

Journal of Medical Microbiology

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A down-flow immunoassay has been developed to detect serum mannan antigens, and the test was recently marketed as the Unimedi Candida monotest. Using 251 serum samples from 105 patients with candidaemia, a comparison of the Unimedi Candida monotest with the Cand-Tec latex agglutination test and 2 microplate enzyme immunoassay tests (Platelia Candida Ag test and Unimedi Candida) was conducted. One hundred and seventy-five febrile patients without clinical and microbiological evidence of fungal infections and pneumocytosis were examined as controls. The Cand-Tec test had a sensitivity of 38 % and a specificity of 82 %. The sensitivity and specificity of the Platelia Candida Ag test, the Unimedi Candida and the Unimedi Candida monotest were 53 and 92 %, 69 and 89 % and 82 and 96 %, respectively. The sensitivity of the Unimedi Candida monotest was significantly (P<0?01) higher than that of the Plateria Candida Ag test for diagnosing candidaemia caused by Candida parapsilosis. The b-D-glucan assay had a high sensitivity of 95 %, with a specificity of 84 %. Of 74 patients with candidaemia whose sera were available before or on positive blood culture sampling, 29 (39 %), 38 (51 %) and 48 (65 %) patients had antigenemia detected using the Platelia Candida Ag test, the Unimedi Candida and the Unimedi Candida monotest, respectively. The Unimedi Candida monotest seems to be a promising tool for the early diagnosis of invasive candidiasis, because the test was sensitive, simple, rapid (approx. 1 h) and cost-effective.

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“…Based on the results of carbohydrate assimilation profiles obtained by ID32C and a Vitek 2 yeast identification system, the isolates were identified as C. famata. For molecular identification, the genomic DNA from the isolates was prepared as described previously (1). The internally transcribed spacer (ITS) region of ribosomal DNA (rDNA) was amplified and sequenced as described previously (2).…”

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confidence: 99%

Candida fermentati as a Cause of Persistent Fungemia in a Preterm Neonate Successfully Treated by Combination Therapy with Amphotericin B and Caspofungin

et al. 2015

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A case of persistent candidemia in a preterm neonate caused by Candida fermentati, identified by sequencing of the internally transcribed spacer region of ribosomal DNA (rDNA), is described. The neonate was treated for 30 days by combination therapy with amphotericin B (AmBisome) and caspofungin with a successful outcome, and no drug-related side effects were observed. CASE REPORTA 27-week-preterm male, weighing 1,040 g, was delivered to a Kuwaiti mother by emergency longitudinal cesarean section in a surgical intensive care unit. On day 0, the baby was diagnosed to have hyaline membrane disease and patent ductus arteriosus with Apgar scores of 7 and 9. Soon after delivery, an umbilical artery catheter (UAC), an umbilical venous catheter (UVC), and a peripheral cannula were placed to provide fluids and medication and the baby was also intubated. On day 5, he was empirically started on ampicillin and amikacin. On day 12, he developed signs and symptoms of sepsis requiring a change in treatment to cloxacillin and amikacin. Since his septic condition continued, the treatment was changed to vancomycin. On day 27, the blood culture grew a yeast (Kw3414/13), which was identified as Candida famata by the use of a Vitek 2 yeast identification (YST ID) card (bioMérieux, Marcy I'Étoile, France). While cultures of UVC and UAC tips were negative for growth, a culture from a long-line tip yielded yeast which was also identified as C. famata. The antifungal drug susceptibility was determined by Etest, and the isolate appeared susceptible to amphotericin B (Table 1). The baby was started on amphotericin B at 1.5 mg/kg of body weight. A blood culture repeated after 2 days of treatment again yielded C. famata, leading to a change of treatment to a lipid formulation of amphotericin B (AmBisome) (8.5 mg once daily). Despite treatment with amphotericin B for 5 days, the blood culture remained positive for C. famata, prompting addition of caspofungin (2.5 mg/kg). Although additional blood cultures performed on day 50, day 53, and day 59 yielded C. famata (identified by the use of a Vitek 2 yeast identification system), there was symptomatic improvement in the condition of the baby. A blood culture repeated on day 66 (after 30 days of combination therapy with amphotericin B and caspofungin) became negative for the yeast. There was no recurrence of infection during 3 weeks of follow-up, and the baby was subsequently discharged in healthy condition.During 6 weeks of follow-up, 6 blood culture isolates were obtained and were characterized. All isolates formed whitish colonies on Sabouraud dextrose agar which became slightly tanned on aging. Microscopic examination revealed ovoid to elongated yeast cells of variable sizes (2 to 5.2 by 2.2 to 5.6 m) occurring singly, in pairs, or in short chains. A Dalmau slide culture on cornmeal agar at 30°C showed well-branched pseudohyphae with clusters of budding yeast cells (Fig. 1). True hyphae were lacking. Growth on ascospore agar and malt extract agar, incubated at 25°C for 30 days, did not show...

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PCR-enzyme immunoassay of rDNA in the diagnosis of candidemia and comparison with amplicon detection by agarose gel electrophoresis

Cited by 43 publications

References 23 publications

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

Evaluation of a newly developed down-flow immunoassay for detection of serum mannan antigens in patients with candidaemia

Candida fermentati as a Cause of Persistent Fungemia in a Preterm Neonate Successfully Treated by Combination Therapy with Amphotericin B and Caspofungin

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