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  • 1
    In: Journal of Medical Microbiology, Microbiology Society, Vol. 72, No. 8 ( 2023-08-25)
    Abstract: Introduction. Invasive mucormycosis (IM) is a potentially fatal infection caused by fungi of the order Mucorales . Histopathology, culture, and radiology are the mainstays of diagnosis, but they are not sufficiently sensitive, resulting in delayed diagnosis and intervention. Recent studies have shown that PCR-based techniques can be a promising way to diagnose IM. Hypothesis/Gap Statement. Early diagnosis of fungal infections using molecular diagnostic techniques can improve patient outcomes, especially in invasive mucormycosis. Aim. The aim of this study was to evaluate the utility of our in-house mould-specific real time PCR assay (qPCR) in comparison with the commercially available real time PCR ( MucorGenius PCR), for the early diagnosis of mucormycosis in tissue samples from patients with suspicion of invasive mucormycosis (IM). This in-house assay can detect and distinguish three clinically relevant mould species, e.g. Aspergillus spp., Mucorales and Fusarium spp. in a single reaction with only one pair of primers, without the need for sequencing. Methodology. We enrolled 313 tissue samples from 193 patients with suspected IM in this prospective study. All cases were classified using EORTC/MSGERC guidelines. All samples were tested using traditional methods, in-house qPCR, and MucorGenius PCR. Results. Using direct microscopy as a gold standard, the overall sensitivity and specificity of in-house qPCR for detection of IM was 92.46% and 80% respectively, while that of the MucorGenius PCR was 66.67% and 90% respectively. However, co-infection of IM and IA adversely affected the performance of MucorGenius PCR in detection of IM. The in-house PCR detected Aspergillus spp. in 14 cases and Fusarium spp . in 4 cases which showed clinical and radiological features of fungal sinusitis. The in-house qPCR also performed better in detecting possible cases of IM. This aids early diagnosis and appropriate treatment to improve patient outcomes. Conclusion. Because the in-house PCR is not only sensitive and specific, but also entirely based on SYBR Green for detection of targets, it is less expensive than probe-based assays and can be used on a regular basis for the diagnosis of IM in resource-constrained settings. It can be used to distinguish between mucormycosis and fungal sinusitis caused by Aspergillus and Fusarium in high-risk patients, as well as to accurately detect Mucorales in fungal co-infection cases.
    Type of Medium: Online Resource
    ISSN: 0022-2615 , 1473-5644
    RVK:
    Language: English
    Publisher: Microbiology Society
    Publication Date: 2023
    detail.hit.zdb_id: 2083944-3
    SSG: 12
    Location Call Number Limitation Availability
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  • 2
    Online Resource
    Online Resource
    Oxford University Press (OUP) ; 2022
    In:  Medical Mycology Vol. 60, No. Supplement_1 ( 2022-09-20)
    In: Medical Mycology, Oxford University Press (OUP), Vol. 60, No. Supplement_1 ( 2022-09-20)
    Abstract:   Background and Objective Molds are emerging as a major cause of life-threatening infections in immunocompromised patients. There is an increasing recognition of the cryptic Aspergillus species, which are organisms that are morphologically indistinguishable yet can be differentiated by molecular methods. These organisms have been known to show a higher minimal inhibitory concentration for the majority of the antifungal agents in vitro. Therefore, correct identification of these cryptic species is very important to administer a proper antifungal agent. In this study, we wish to identify and characterize the cryptic species of Aspergillus from all clinical samples. Methods Patient routine samples like broncho alveolar lavage, endo-tracheal aspirate, sputum, pus, tissue, and CSF from various wards, OPDs, and ICUs of All India Institute of Medical Sciences, New Delhi which were processed in the Mycology laboratory, department of Microbiology showing growth of Aspergillus species were included in this study. Identification of the isolates was done using phenotypic methods and by Matrix Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF). For the analysis of cryptic species, the isolates underwent PCR and then sequencing of the ß-tubulin gene was done. Antifungal susceptibility testing was done using micro broth dilution as per the CLSI method. Results Of the 92 isolates, using morphological methods, 53 were identified as A. fumigatus, 33 were identified as A. flavus, 3 as A. nidulans, 2 as A. terreus, and 1 as A. niger MALDI-TOF (Vitek MS database) misidentified 2 isolates of A. nidulans and 1 isolate of A, stellatus as A. fumigatus and 1 isolate of A. tubingensis as A. niger. The ß-tubulin sequence analysis for the identification of cryptic species revealed that 2 isolates (2.08%) were cryptic, one was A. stellatus morphologically identified as A. nidulans and another one was A. tubingensis morphologically identified as A. niger. Conclusion Currently the available data on cryptic Aspergillus species is very limited. Because of its varied susceptibility pattern, it is important to identify the Aspergillus isolates to its species level. There is also a need for expansion of the number of strains for each species in MALDI-TOF MS databases for convenient, faster, and correct identification.
    Type of Medium: Online Resource
    ISSN: 1369-3786 , 1460-2709
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2022
    detail.hit.zdb_id: 2020733-5
    SSG: 12
    Location Call Number Limitation Availability
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  • 3
    In: Journal of Medical Microbiology, Microbiology Society, Vol. 72, No. 9 ( 2023-09-15)
    Type of Medium: Online Resource
    ISSN: 0022-2615 , 1473-5644
    RVK:
    Language: English
    Publisher: Microbiology Society
    Publication Date: 2023
    detail.hit.zdb_id: 2083944-3
    SSG: 12
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
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