Quantitative Polymerase Chain Reaction Detection of Circulating DNA in Serum for Early Diagnosis of Mucormycosis in Immunocompromised Patients

Millon, Laurence; Larosa, Fabrice; Lepiller, Quentin; Legrand, Faézeh; Rocchi, Steffi; Daguindau, Étienne; Scherer, Émeline; Bellanger, Anne‐Pauline; Leroy, J.; Grenouillet, Frédéric

doi:10.1093/cid/cit094

Cited by 187 publications

(137 citation statements)

References 26 publications

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“…in serum samples, can guide treatment effectively and improve patients' outcomes. Furthermore, other real-time assays detect either a single Mucorales genus (Bernal-Martínez et al, 2013;Lengerova et al, 2014) or only groups of Mucorales pathogens, missing other clinically relevant species (Millon et al, 2013;Hata et al, 2008). In detail, Voigt et al (1999) used taxon-specific primer pairs in an end point PCR which would require performing 13 different assays.…”

Section: Discussionmentioning

confidence: 99%

“…Bernal-Martínez et al (2013) used a multiplex qPCR assay designed to detect only the genera Rhizopus and Mucor. Also very specific assays were used by Millon et al (2013). Three real-time assays are necessary to detect the four different genera (Lichtheimia, Rhizopus, Rhizomucor and Mucor).…”

Section: Discussionmentioning

confidence: 99%

“…They are faster and more straightforward than end point PCR assays, with a lower risk of contamination. In the future, these tests might also be used as a screening tool, including the analysis of blood specimens (Millon et al, 2013). However, currently, inter-laboratory standardization seems to be difficult since (i) Mucorales contains a large variety of different fungal species, (ii) Mucorales infections occur with a relatively low incidence (but are probably underestimated) and therefore few specimens are collected, (iii) different clinical material is used for the detection of Mucorales, and (iv) different poorly standardized in-house protocols are used for DNA extraction and PCR amplification.…”

Section: Discussionmentioning

confidence: 99%

“…There are reports using different sources for detecting Mucorales DNA from fresh or formalin-fixed paraffin-embedded tissue (FF-PET), bronchoalveolar lavages or serum samples (Bialek et al, 2005;Rickerts et al, 2006;Kasai et al, 2008;Millon et al, 2013). Also, different methods to detect Mucorales by PCR have been described, including conventional, semi-nested and real-time (DNA intercalating dyes, molecular-beacon or hydrolysis probe-based) PCR (Millon et al, 2013;Bialek et al, 2005;Voigt et al, 1999;Hata et al, 2008;Bernal-Martínez et al, 2013;Hrncirova et al, 2010;Lengerova et al, 2014); each suffers from different disadvantages such as high turnaround time, vulnerability to contamination or limited detection of selected species or genera.…”

Section: Introductionmentioning

confidence: 99%

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Development and application of two independent real-time PCR assays to detect clinically relevant Mucorales species

Springer

Goldenberger

Schmidt

et al. 2016

Journal of Medical Microbiology

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PCR-based detection of Mucorales species could improve diagnosis of suspected invasive fungal infection, leading to a better patient outcome. This study describes two independent probe-based real-time PCR tests for detection of clinically relevant Mucorales, targeting specific fragments of the 18S and the 28S rRNA genes. Both assays have a short turnaround time, allow fast, specific and very sensitive detection of clinically relevant Mucorales and have the potential to be used as quantitative tests. They were validated on various clinical samples (fresh and formalin-fixed paraffin-embedded specimens, mainly biopsies, n517). The assays should be used as add-on tools to complement standard techniques; a combined approach of both real-time PCR assays has 100 % sensitivity. Genus identification by subsequent sequencing is possible for amplicons of the 18S PCR assay. In conclusion, combination of the two independent Mucorales assays described in this study, 18S and 28S, detected all clinical samples associated with proven Mucorales infection (n510). Reliable and specific identification of Mucorales is a prerequisite for successful antifungal therapy as these fungi show intrinsic resistance to voriconazole and caspofungin.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Development and application of two independent real-time PCR assays to detect clinically relevant Mucorales species

Springer

Goldenberger

Schmidt

et al. 2016

Journal of Medical Microbiology

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“…Millon et al (15) recently published a strategy based on monitoring the mucormycete DNA in serum samples from patients with histopathologically proven IM. Testing serum samples is very convenient, but due to the low amount of circulating fungal DNA, the positivity of serum samples can be expected only in advanced stages of the disease.…”

Section: Discussionmentioning

confidence: 99%

Rapid Detection and Identification of Mucormycetes in Bronchoalveolar Lavage Samples from Immunocompromised Patients with Pulmonary Infiltrates by Use of High-Resolution Melt Analysis

et al. 2014

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e Rapid differential diagnostics of pulmonary infiltrates suspected of invasive fungal disease in an immunocompromised host and early initiation of effective antifungal therapy are crucial for patient outcomes. There are no serological tests available to detect mucormycetes; therefore, PCR-based methods are highly suitable. We validated our previously published PCR followed by highresolution melt analysis (PCR/HRMA) to detect Rhizopus spp., Rhizomucor pusillus, Lichtheimia corymbifera, and Mucor spp. in bronchoalveolar lavage (BAL) samples from immunocompromised patients who were at risk of invasive fungal disease. All PCR/ HRMA-positive samples were retested using novel real-time quantitative PCR (RQ PCR) assays specific to the species identified. In total, between January 2009 and December 2012 we analyzed 99 BAL samples from 86 patients with pulmonary abnormalities using PCR/HRMA. Ninety (91%) BAL samples were negative, and 9 (9%) samples were positive. The sensitivity and specificity of PCR/HRMA were 100% and 93%, respectively. By combining the positive results of PCR/HRMA with positive RQ PCR results, the specificity was raised to 98%. PCR/HRMA, due to its high negative predictive value (99%), represents a fast and reliable tool for routine BAL sample screening for the differential diagnosis of pulmonary infiltrates in immunocompromised patients for the four most clinically important mucormycetes.

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A successfully treated gastric mucormycosis in an immunocompetent patient: Case report and literature review

Albtoosh,

Shaf'ei,

Al Hayek

et al. 2023

Clinical Case Reports

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Mucormycosis is an opportunistic fungal infection that usually affects patients with diabetes mellitus or immunosuppression. The fungus invades the nearby blood vessels leading to thrombosis and necrosis of the organs involved. Although Mucorales can invade any organ in the body, the gastrointestinal system is an uncommon site for infection. Mucormycosis is a fatal infection, and prompt intervention is required to ensure survival. In this report, we present a case of a 46-year-old man with history of valve replacement surgery on warfarin, who admitted with abdominal pain and life-threatening gastrointestinal bleeding.Esophagogastroduodenoscopy revealed an active gastric ulcer bleeding, and the diagnosis of mucormycosis infection was confirmed with direct microscopy and histopathological evaluation from a tissue biopsy. Typically, antifungal therapy alone is inadequate to control mucormycosis infection and surgical intervention is often required. Our patient was successfully treated using antifungal therapy alone. This report presents a rare case of gastrointestinal mucormycosis in setting of valve replacement and was successfully treated with antifungal therapy.

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Quantitative Polymerase Chain Reaction Detection of Circulating DNA in Serum for Early Diagnosis of Mucormycosis in Immunocompromised Patients

Cited by 187 publications

References 26 publications

Development and application of two independent real-time PCR assays to detect clinically relevant Mucorales species

Development and application of two independent real-time PCR assays to detect clinically relevant Mucorales species

Rapid Detection and Identification of Mucormycetes in Bronchoalveolar Lavage Samples from Immunocompromised Patients with Pulmonary Infiltrates by Use of High-Resolution Melt Analysis

A successfully treated gastric mucormycosis in an immunocompetent patient: Case report and literature review

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