Usefulness of molecular biology performed with formaldehyde-fixed paraffin embedded tissue for the diagnosis of combined pulmonary invasive mucormycosis and aspergillosis in an immunocompromised patient

Hofman, Véronique; Dhouibi, Abdelmajid; Butori, Catherine; Padovani, B.; Gari-Toussaint, M.; García-Hermoso, Dea; Baumann, Michèle; Vénissac, Nicolas; Cathomas, Gieri; Hofman, Paul

doi:10.1186/1746-1596-5-1

Cited by 59 publications

(40 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To date, PCR-based molecular identification methods, including PCR (1,4), seminested PCR (19), quantitative realtime PCR (3), multiplex PCR (12), PCR-restriction fragment length polymorphism (RFLP) (9), randomly amplified polymorphic DNA (RAPD) (20), and ITS sequence analysis (2,22), have been used to identify Aspergillus and Mucorales species. However, these assays are not readily adaptable for use in the clinical lab due to disadvantages, including the following: (i) PCR, seminested PCR, and real-time PCR can identify only one species at a time, (ii) it is difficult to draw clear conclusions from the results of RFLP and RAPD, (iii) the number of primers for the Multiplex PCR system is limited, and (iv) sequence analysis is time-consuming and often fails to identify samples with mixed infections and samples of poor quality or that are in small quantities.…”

Section: Discussionmentioning

confidence: 99%

“…The T m values for oligonucleotide primers or probes were provided by the company that synthesized the oligonucleotides (Sangon). c These primer and probe sequences have been previously published, but some of them were modified for this study (4,8,15,19). All of the other probes were designed specifically for this study.…”

Section: Detection and Identification Of Fungal Strains By Rlb Assaymentioning

confidence: 99%

See 1 more Smart Citation

Simultaneous Detection and Identification of Aspergillus and Mucorales Species in Tissues Collected from Patients with Fungal Rhinosinusitis

Zhao

Wan

et al. 2011

J Clin Microbiol

View full text Add to dashboard Cite

Rapid detection and differentiation of Aspergillus and Mucorales species in fungal rhinosinusitis diagnosis are desirable, since the clinical management and prognosis associated with the two taxa are fundamentally different. We describe an assay based on a combination of broad-range PCR amplification and reverse line blot hybridization (PCR/RLB) to detect and differentiate the pathogens causing fungal rhinosinusitis, which include five Aspergillus species (A. fumigatus, A. flavus, A. niger, A. terreus, and A. nidulans) and seven Mucorales species (Mucor heimalis, Mucor racemosus, Mucor cercinelloidea, Rhizopus arrhizus, Rhizopus microsporus, Rhizomucor pusillus, and Absidia corymbifera). The assay was validated with 98 well-characterized clinical isolates and 41 clinical tissue specimens. PCR/RLB showed high sensitivity and specificity, with 100% correct identifications of 98 clinical isolates and no cross-hybridization between the species-specific probes. Results for five control isolates, Candida albicans, Fusarium solani, Scedosporium apiospermum, Penicillium marneffei, and Exophiala verrucosa, were negative as judged by PCR/RLB. The analytical sensitivity of PCR/RLB was found to be 1.8 ؋ 10 Rhinosinusitis is a common disease with multiple manifestations and can even cause death in acute invasive cases. Fungi are considered to be major etiological agents, of which members of the order Mucorales and genus Aspergillus (order Eurotiales) are the most pathogenic (5, 10, 13). The course, therapeutic treatment, and prognosis of fungal rhinosinusitis caused by Aspergillus and Mucorales species are radically different; therefore, early diagnosis and accurate identification of pathogenic fungal species are crucial for effective treatment and clinical decision-making (21). Currently, diagnosis of fungal sinusitis still depends on histopathological examination and culture from nasal biopsy, but conventional culture-based phenotypic identification techniques often include significant delays and can fail to yield growth in clinical samples (17). In a significant number of cases, fungal culture is negative and only formalin-fixed paraffin-embedded (PE) tissue specimens are available for diagnosis of fungal infection. However, rapid diagnosis of surgical tissues is urgently needed in acute invasive infection cases. In addition, histopathological observations of fungal shape and arrangement may not be sufficient for the accurate identification of fungal species if only a limited quantity of anamorphic fungal hyphae is present. Therefore, to improve the outcome for fungal rhinosinusitis patients, the rapid and accurate detection and identification of pathogenic fungal species are needed to allow early initiation of targeted therapy.In this study, we developed an assay combining broad-range PCR amplification and reverse line blot hybridization (PCR/ RLB) to detect and differentiate Aspergillus and Mucorales pathogens in tissue specimens. The fungal pathogens tested included five Aspergillus species (Aspergillus fumigatus, A. f...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Detection and Identification Of Fungal Strains By Rlb Assaymentioning

confidence: 99%

Simultaneous Detection and Identification of Aspergillus and Mucorales Species in Tissues Collected from Patients with Fungal Rhinosinusitis

Zhao

Wan

et al. 2011

J Clin Microbiol

View full text Add to dashboard Cite

show abstract

“…and Candida or mucoraceous genera have been described and pose important diagnostic dilemmas. To be able to identify dual infections, it is crucial to have alternative diagnostic testing of tissues such as immunohistochemistry, in situ hybridization, or PCR, which at this time are available only for research use (70,147). In addition, in invasive pulmonary aspergillosis, cultures are positive in only 50% of bronchoalveolar lavage fluid specimens (148), and organisms recovered from BAL fluid samples may reflect colonization rather than the actual pathogen.…”

Section: Diseases Where Hyphae Are Usually Seen In Tissuementioning

confidence: 99%

“…Poor staining of hyphae with GMS should suggest mucormycosis. To be able to specifically identify Mucorales in tissues or to detect dual infections by Mucorales genera and other fungi, it is important to use immunohistochemistry, in situ hybridization, or PCR (70,147).…”

Section: Mucormycosis (Zygomycosis) (I) Epidemiologicmentioning

confidence: 99%

Histopathologic Diagnosis of Fungal Infections in the 21st Century

2011

View full text Add to dashboard Cite

SUMMARY Fungal infections are becoming more frequent because of expansion of at-risk populations and the use of treatment modalities that permit longer survival of these patients. Because histopathologic examination of tissues detects fungal invasion of tissues and vessels as well as the host reaction to the fungus, it is and will remain an important tool to define the diagnostic significance of positive culture isolates or results from PCR testing. However, there are very few instances where the morphological characteristics of fungi are specific. Therefore, histopathologic diagnosis should be primarily descriptive of the fungus and should include the presence or absence of tissue invasion and the host reaction to the infection. The pathology report should also include a comment stating the most frequent fungi associated with that morphology as well as other possible fungi and parasites that should be considered in the differential diagnosis. Alternate techniques have been used to determine the specific agent present in the histopathologic specimen, including immunohistochemistry, in situ hybridization, and PCR. In addition, techniques such as laser microdissection will be useful to detect the now more frequently recognized dual fungal infections and the local environment in which this phenomenon occurs.

show abstract

“…was first identified during the course of the disease by cytological examination and culture (A. fumigatus) of bronchoalveolar fluid. Hyphae of Mucorales (Rhizopus microsporus) were subsequently identified by culture of a tissue specimen taken from the left inferior pulmonary lobe, which was surgically resected two days before the patient died (11).…”

Section: Işıktaş Sayılar E Et Al: Posttransplant Invasive Fungal Infementioning

confidence: 99%

Mixed Fungal Infection in Early Period after Kidney Transplantation: A Case Report

Sayilar¹,

Ayar²,

Akalın³

et al. 2015

Turk Neph Dial Transpl

View full text Add to dashboard Cite

Invasive fungal infections have a rapid and frequently fatal course in patients with solid organ transplantations. Mostly Aspergillus spp., Mucorales spp., Candida spp. and Cryptococcus neoformans are causal pathogens for opportunistic infections. Aspergillus spp. and Mucorales spp. especially lead to invasive fungal infections at rhino-cerebral area; they show similar radiological and clinical signs and they lead to invasive fungal co-infections. In case of any doubt about invasive fungal infections, antifungal treatment should be initiated as soon as possible and immunosuppressive treatment should be considered. This case presentation is about a patient at 51 years of age who developed invasive rhinocerebral mixed fungal infection in 4 weeks following renal transplant.kEy worDS: Kidney transplant, Aspergillus, Mucor, Sinusitis ÖzSolid organ transplantasyonu yapılan hastalarda invaziv fungal infeksiyonlar hızlı ve çoğu zaman ölümcül seyir göstermektedir. Çoğunlukla Aspergillus, Mucor, Candida, ve Cryptococcus neoformans oppurtunistik enfeksiyon etkenleridir. Özellikle Aspergillus ve Mucor rinoserebral bölgede invaziv fungal enfeksiyona yol açarlar, klinik ve radyolojik olarak benzer bulgular gösterirler ve bazen de bir arada invaziv enfeksiyona yol açarlar. İnvaziv funfal infeksiyondan şüphelenildiğinde mümkün olan en kısa sürede antifungal tedavi başlanmalı ve immünsupresif tedavi gözden geçirilmelidir. Bu olgu sunumunda 51 yaşında, renal transplant sonrası 4. haftada invaziv rinoserebral fungal enfeksiyon gelişen bir hasta sunulmuştur.anahtar SözCüklEr: Böbrek nakli, Aspergillus, Mucor, Sinüzit

show abstract

Usefulness of molecular biology performed with formaldehyde-fixed paraffin embedded tissue for the diagnosis of combined pulmonary invasive mucormycosis and aspergillosis in an immunocompromised patient

Cited by 59 publications

References 45 publications

Simultaneous Detection and Identification of Aspergillus and Mucorales Species in Tissues Collected from Patients with Fungal Rhinosinusitis

Simultaneous Detection and Identification of Aspergillus and Mucorales Species in Tissues Collected from Patients with Fungal Rhinosinusitis

Histopathologic Diagnosis of Fungal Infections in the 21st Century

Mixed Fungal Infection in Early Period after Kidney Transplantation: A Case Report

Contact Info

Product

Resources

About