Nested PCR for the detection of Aspergillus species in maxillary sinus samples of patients with chronic sinusitis

Małek, Marianna; Bogusz, Bożena; Mrowiec, Paulina; Szuta, Mariusz; Opach, Maciej; Skiba‐Kurek, Iwona; Nowak, Paweł; Klesiewicz, Karolina; Budak, Alicja; Karczewska, E

doi:10.1016/j.riam.2018.04.001

Cited by 4 publications

(4 citation statements)

References 25 publications

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“…In support of this decision, Małek et al 19 reported that PCR showed perfect concordance with histology with sensitivity and PPV of 90% and specificity and NPV of 98.3%, but additionally, it diagnosed a sample showed growth of Asp. fumigatus despite the negative histological examination.…”

Section: Discussionmentioning

confidence: 92%

Nested Polymerase Chain Reaction versus Enzyme Linked Immunosorbent Assay for Rapid Diagnosis of Sinus Aspergillosis

Saeed

Fouad

Elshaboury

2019

Egyptian Journal of Medical Microbiology

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Fungal infections of the nose and paranasal sinuses are classified into invasive and non invasive types. Rapid detection and identification of causative fungi is crucial for effective treatment and avoidance of complications. Objectives: To evaluate diagnostic outcome of ELISA detection of galactomannan (GM) in sinus lavage aspirate as a rapid minimally invasive diagnostic test for Aspergillus spp. sinus infection in comparison to fungus DNA detection by nested PCR. Methodology: 95 chronic rhinosinusitis (CRS) patients had manifestations since ≥12 weeks despite of medical therapy were clinically and radiologically evaluated and underwent sinus lavage. Lavage aspirate was used for microbiological examination, PCR testing for Aspergillus DNA and ELISA detection of GM. Results: PCR defined Aspergillus DNA in 56 samples, culture defined 53 positive samples for Aspergillus spp. infection and ELISA detected GM antigen in 49 samples. In comparison to PCR result, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) and accuracy rates of culture were 73.2%, 69.3%, 77.4%, 64.3% and 71.6% while ELISA detected GM antigen were 84. 6%, 88.4%, 89.8%, 82.6% and 86.3%, respectively. ELISA results significantly correlated with results of culture and PCR with a positive significant correlation between PCR and culture results. ROC curve analysis defined ELISA detection of GM as more significant predictor for result of culture (AUC =0.792, p<0.001) than PCR test (AUC=0.708, p=0.001). Regression analysis defined PCR testing as the significant predictor for culture results (β=0.529, p<0.001).Conclusion: Fungal CRS causes resistance to conventional treatment and must be investigated before surgical-decision taking. Preoperative ELISA determination of GM antigen could identify patients with aspergillosis sinus infection with high accuracy and specificity. For suspicious cases especially if negative for GM, nested PCR could approach the diagnosis.

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Section: Discussionmentioning

confidence: 92%

Nested Polymerase Chain Reaction versus Enzyme Linked Immunosorbent Assay for Rapid Diagnosis of Sinus Aspergillosis

Saeed

Fouad

Elshaboury

2019

Egyptian Journal of Medical Microbiology

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“…Despite extensive studies, there are still serious controversial topics about the best way to diagnose IFIs, especially when IA is suspected in patients undergoing chemotherapy or BMT (18). Given the low sensitivity of mycological culture to detect aspergillosis, relying on culture alone is not enough to rule out the infection and even new methods such as polymerase chain reaction (PCR) are not superior to cultures for detecting invasive mold infections (18,20). IA in the patients with hematologic malignancies usually causes sinopulmonary involvement because their immunocompromised status may provide an opportunity for growing spores and invade sinonasal mucosal tissue.…”

Section: Discussionmentioning

confidence: 99%

Role of Nasal Lavage Fluid Galactomannan Levels For Diagnosis of Invasive Aspergillosis in Patients with Leukemia

et al. 2021

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Background: Invasive aspergillosis (IA) is the most prevalent invasive infection with high mortality among patients with leukemia. Early diagnosis of IA has been a challenging topic in this group of patients. Objectives: In this study, we evaluated the galactomannan levels of nasal lavage fluid (NALF) as a possible auxiliary method for IA diagnosis in patients with leukemia. Methods: In a prospective study, 32 adult patients with leukemia who were taking induction and/or consolidation chemotherapy with fever and neutropenia were included. In all patients, galactomannan (GM) levels of serum and NALF, and mycological examinations were evaluated before the first dose of antifungal therapy. Results: Fourteen (43.7%) patients had NALF GM ≥ 0.5; however, in 16 (50%) patients the level of serum GM was ≥ 0.5. The elevated level of NALF GM had a significant association with the proven IA cases (P = 0.048). The GM level of NALF with a cut-off value of 0.45 (by receiver operating characteristic curve analysis) had 78% sensitivity and 64% specificity for the diagnosis of invasive aspergillosis (P = 0.033). Conclusions: Due to its non-invasive nature, GM level of NALF may be contributory to be used as part of the diagnostic work‐up of IA particularly in leukemic patients with thrombocytopenia which prohibits acquiring bronchoalveolar lavage.

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“…A variety of PCR-based methods, including nested PCR [10,11], PCR-RFLP [12], SYBR green-based quantitative PCR [13], PCR-sequencing [14], and real-time PCR [15][16][17], have been introduced to directly detect/identify fungi from clinical samples. Among the mentioned methods, multiplex PCR has the advantage of being able to detect/identify several genera and species of fungi simultaneously in the shortest time possible in a PCR reaction [9].…”

Section: Introductionmentioning

confidence: 99%

Molecular strategy for the direct detection and identification of the most common fungal community in cerumen specimens by multiplex PCR

Jabrodini,

Sohrabizdeh,

Aboutalebian

et al. 2023

Journal of Medical Microbiology

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Introduction. Otomycosis is a superficial fungal infection that is responsible for approximately 9–27 % of otitis externa. However, fungal communities in otomycosis are varied, but Aspergillus spp. and Candida spp. are the most common causes of this infection. Hypothesis Statement. The multiplex PCR assay is postulated to be able to directly detect more than one fungal genus in cerumen specimens. Aim. This study aimed to develop and evaluate the role of the multiplex PCR assay in detecting the most common genus of fungi that cause otomycosis directly from the cerumen specimens. Methodology. To detect Candida and Aspergillus/Penicillium genera, three pairs of primers, including pan-fungal, pan-Candida, and pan-Aspergillus/Penicillium, were used in a multiplex PCR. In order to evaluate the performance and reproducibility of the multiplex PCR. the cerumen of 140 patients suspected of otomycosis were investigated. Results. Pan-Candida and pan-Aspergillus/Penicillium primers were designed to amplify the ITS1–5.8S–ITS2 region and the β-tubulin gene, respectively. In the multiplex PCR assay, 64 (47.40 %) and 118 (87.40 %) specimens were positive with pan-Candida and pan-Aspergillus/Penicillium primers, respectively. Double amplicon bands of Candida and Aspergillus were obtained in 51 (37.77 %) specimens. In the culture method, yeast (n=18, 13.33 %) and mould (n=117, 86.66 %) were isolated from 135 cerumen specimens. The sensitivity, specificity, and positive and negative predictive values of the multiplex PCR assays using culture method results as the gold standard were determined to be 94, 33, 97, and 22 %, respectively. Conclusion. In our study, multiplex PCR assays enabled simultaneous detection of two common genera of the causative agent of otomycosis in a cerumen specimen. Regarding the high sensitivity of the first step of the multiplex PCR assay, this assay may be used for the direct detection of Candida and Aspergillus genera in other clinical specimens.

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Nested PCR for the detection of Aspergillus species in maxillary sinus samples of patients with chronic sinusitis

Cited by 4 publications

References 25 publications

Nested Polymerase Chain Reaction versus Enzyme Linked Immunosorbent Assay for Rapid Diagnosis of Sinus Aspergillosis

Nested Polymerase Chain Reaction versus Enzyme Linked Immunosorbent Assay for Rapid Diagnosis of Sinus Aspergillosis

Role of Nasal Lavage Fluid Galactomannan Levels For Diagnosis of Invasive Aspergillosis in Patients with Leukemia

Molecular strategy for the direct detection and identification of the most common fungal community in cerumen specimens by multiplex PCR

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