Clinical utility of a panfungal polymerase chain reaction assay for invasive fungal diseases in patients with haematologic disorders

Sugawara, Yumiko; Nakase, Kazunori; Nakamura, Akiko; Ohishi, Kohshi; Sugimoto, Yuka; Fujieda, Atushi; Monma, Fumihiko; Suzuki, Kei; Masuya, Masahiro; Matsushima, Yuta; Wada, Hideo; Nobori, Tsutomu; Katayama, Naoyuki

doi:10.1111/ejh.12078

Cited by 44 publications

(37 citation statements)

References 45 publications

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“…1). Twentythree studies provided data for PCR based on the 2002 EORTC/ MSG criteria , and 14 did so by using the revised (2008) criteria (47)(48)(49)(50)(51)(52)(53)(54)(55)(56)(57)(58)(59)(60) (Table 1).…”

Section: Resultsmentioning

confidence: 99%

PCR in Diagnosis of Invasive Aspergillosis: a Meta-Analysis of Diagnostic Performance

et al. 2014

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Invasive aspergillosis is a difficult-to-diagnose infection with a high mortality rate that affects high-risk groups such as patients with neutropenia and hematologic malignancies. We performed a bivariate meta-analysis of diagnostic data for an Aspergillus sp. PCR assay with blood specimens from high-risk hematology patients. We included all studies involving human subjects that assessed the performance of any PCR assay for invasive aspergillosis in whole blood or serum and that used the European Organization for the treatment of Cancer/Mycoses Study Group criteria as a reference standard. Three investigators independently searched the literature for eligible studies and extracted the data. Out of a total of 37 studies, 25 met strict quality criteria and were included in our evidence synthesis. Twenty-five studies with 2,595 patients were analyzed. The pooled diagnostic performance of whole-blood and serum PCR assays was moderate, with a sensitivity and specificity of 84% (95% confidence interval [CI], 75 to 91%) and 76% (95% CI, 65 to 84%), respectively, suggesting that a positive or negative result is unable, on its own, to confirm or exclude a suspected infection. The performance of a PCR assay of serum was not significantly different from that of whole blood. Notably, at least two positive PCR test results were found to have a specificity of 95% and a sensitivity of 64% for invasive infection, achieving a high positive likelihood ratio of 12.8. Importantly, the European Aspergillus PCR Initiative (EAP-CRI) recommendations improved the performance of the PCR even further when at least two positive specimens were used to define PCR positivity. In conclusion, two positive PCR results should be considered highly indicative of an active Aspergillus sp. infection. Use of the EAPCRI recommendations by clinical laboratories can further enhance PCR performance. D espite advances in treatment and supportive care, invasive aspergillosis (IA) is associated with significant morbidity and mortality rates, especially among patients with hematologic malignancies and hematopoietic stem cell transplant (HSCT) recipients (1, 2). Systemic antifungal prophylaxis is widely used in this patient population (3, 4), and patients with recurrent or persistent fever and prolonged neutropenia frequently require empirical coverage with antifungal agents (5). Timely and accurate diagnosis of an active infection is needed in order to initiate targeted antifungal therapy and avoid unnecessary antifungal treatment, which is often accompanied by a multitude of side effects and the cumulative risk of resistance.There is a need for the development of newer diagnostic techniques that would ideally be able to identify IA rapidly, noninvasively, and at an early stage. To aid in this endeavor, the European Organization for the treatment of Cancer/Mycoses Study Group (EORTC/MSG) developed specific criteria for the diagnosis of IA in 2002 (6), which were later revised in 2008 (7), in an attempt to provide the elusive "gold standard" to which any newe...

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Section: Resultsmentioning

confidence: 99%

PCR in Diagnosis of Invasive Aspergillosis: a Meta-Analysis of Diagnostic Performance

et al. 2014

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“…(235), and Fusarium spp. (236), but the possibility for clinical implementation of these assays seems unlikely for the foreseeable future, either because serologic assays are already highly sensitive and specific, thus lowering the need for molecular techniques, as is the case for Cryptococcus spp. and Coccidioides spp.…”

Section: Pcrmentioning

confidence: 99%

“…In such cases, PCR facilitates species identification, which cannot be achieved through microscopy but can serve an important role in guiding antifungal therapy. Multiplex PCR has also been tested as a method to detect fungal species in whole-blood (236,238,(243)(244)(245), serum (246), or BAL fluid (247) samples from patients at high risk for IFIs. The results are variable, but most studies report superior sensitivities and specificities of Ͼ80% (236,238,243,248).…”

Section: Pcrmentioning

confidence: 99%

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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“…Between January 1980 and September 1989, 87 cases of dermal and ocular Fusarium infection were diagnosed in the CIB (20) Identification of the species level of fungi belonging to Fusarium using classical methods is difficult. It is time consuming and it has limitations in terms of sensitivity and specificity (34,35). On the other hand, Fusarium taxonomy complexity is widely known (18,31,(36)(37)(38)(39)(40)(41).…”

Section: Discussionmentioning

confidence: 99%

Molecular identification of clinical isolates of Fusarium in Colombia

et al. 2018

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Objective Identifying Fusarium isolates from mycosis symptomatic patients through molecular techniques as PCR and sequencing. Methods In this study, samples were taken from 101 mycosis symptomatic patients in-between [2004][2005][2006]. To determine isolates belonging to the Fusarium genus, the DNAr 28S region was amplified through PCR and specific PCR primers further confirmed their identity to the species level. Additionally, in order to confirm the identity of the species of the isolates, 75 isolates of these were analyzed by partial sequencing of the 28S rDNA and the TEF1-α gene. Results The 28S rDNA portion detected all 101 isolates as belonging to Fusarium and the PCR specific primers detected 52 and 29 isolates as F. oxysporum and F. solani, respectively; 34 and 41 of these, afterwards studied by partial sequencing of the 28S rDNA and TEF1-α genes respectively, were effectively identified by the technique. Conclusion From all the molecular markers used to identify Fusarium isolates, the sequence of the TEF1-α gene provided the best resolution in the identification of species level; however it is possible to discriminate between F. oxysporum and F. solani isolates by PCR, in most of the cases, what is important considering the simplicity of the technique and a faster diagnosis.Key Words: PCR; rDNA; sequence analysis; elongation factor (source: MeSH, NLM). RESUMENObjetivo Identificar aislamientos de Fusarium en pacientes con micosis por medio de las técnicas moleculares de PCR y secuenciación. Métodos Se tomaron 101 muestras de pacientes con micosis sintomática, entre los años 2004 y 2006. Para la detección de aislamientos como pertenecientes al género Fusarium, se amplificó parcialmente por PCR la región 28S del DNAr; y posteriormente -para la detección de la especie de Fusarium-se utilizaron cebadores específicos para F. oxysporum y F. solani. La verificación de la identidad de la especie de los aislamientos se hizo por secuenciación parcial de los genes 28S DNAr y TEF1-α. Resultados El total de 101 aislamientos fueron detectados como pertenecientes al género Fusarium utilizando un cebador universal de la region 28S DNAr; 52 y 29 aislamientos se detectaron como F. oxysporum o F. salani, respectivamente con los cebadores específicos, y la secuenciación parcial de los genes 28S rDNA o TEF1-α confirmó la identidad de las especies. Conclusión La secuenciación parcial del gen TEF1-α es aún el mejor marcador molecular para identificar aislamientos de Fusarium a nivel de especie. Sin embargo, en la mayoría de los casos es posible discriminar entre aislamientos de F. oxysporum y F. solani por PCR con cebadores específicos, lo que proporciona una ventaja importante considerando la simplicidad de la técnica y el rápido diagnóstico.Palabras Clave: ADN ribosómico; PCR; secuenciación; elongación (fuente: DeCS, BIREME).

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Clinical utility of a panfungal polymerase chain reaction assay for invasive fungal diseases in patients with haematologic disorders

Cited by 44 publications

References 45 publications

PCR in Diagnosis of Invasive Aspergillosis: a Meta-Analysis of Diagnostic Performance

PCR in Diagnosis of Invasive Aspergillosis: a Meta-Analysis of Diagnostic Performance

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

Molecular identification of clinical isolates of Fusarium in Colombia

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