Utility of in-house and commercial PCR assay in diagnosis of Covid-19 associated mucormycosis in an emergency setting in a tertiary care center

Author:

Pandey Mragnayani1,Sachdev Janya1,Yadav Renu Kumari1,Sharad Neha2,Kanodia Anupam3,Biswas Jaya1,Janani R. Sruti1,Gupta Sonakshi1,Singh Gagandeep1,Ekka Meera4,Rana Bhaskar1,Gourav Sudesh1,Thakar Alok3,Biswas Ashutosh5,Sikka Kapil3,Mathur Purva2,Pushker Neelam6,Jyotsna Viveka P.7,Kumar Rakesh3,Soneja Manish5,Wig Naveet5,Srivastava M. V. Padma8,Xess Immaculata1

Affiliation:

1. Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India

2. Department of Lab medicine JPNATC, All India Institute of Medical Sciences, New Delhi, India

3. Department of ENT, All India Institute of Medical Sciences, New Delhi, India

4. Department of Emergency Medicine, All India Institute of Medical Sciences, New Delhi, India

5. Department of Medicine, All India Institute of Medical Sciences, New Delhi, India

6. Department of Ophthalmology, All India Institute of Medical Sciences, New Delhi, India

7. Department of Endocrinology, All India Institute of Medical Sciences, New Delhi, India

8. Department of Neurology, All India Institute of Medical Sciences, New Delhi, India

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.

Funder

All-India Institute of Medical Sciences

Publisher

Microbiology Society

Subject

Microbiology (medical),General Medicine,Microbiology

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