Diagnostic Performance and Clinical Utility of Conventional PCR Assay in Early Diagnosis of COVID-19 Associated Rhino-Orbito-Cerebral Mucormycosis
Abstract
:1. Introduction
2. Methods
2.1. Study Design
2.2. Clinical Data of Patients and Staging
2.3. Surgical Intervention and Sample Collection
2.4. Conventional Microbiology Examination
2.5. Standardisation of PCR Assay
2.6. Analytical Sensitivity and Specificity of Primers
2.7. Nucleotide Sequencing and Homology Analysis
2.8. Data Compilation and Statistical Analysis
3. Results
3.1. Demographic and Clinical Details
3.2. Diagnostic Performance of KOH + CFW Smear
3.3. Diagnostic Performance of Culture
3.4. Diagnostic Performance of PCR Assay
3.5. Nucleotide Sequencing
3.6. Efficacy of Nasal Biopsy and Endonasal Swab in Detection of Fungal Pathogens
3.7. Comparison of the Diagnostic Performance of Various Tests
3.8. Treatment Outcomes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Disease Severity | ROC Gradng | Grade | Clinical + Nasal Endoscopy + Imaging Findings | No of Patients (n = 33) (%) |
---|---|---|---|---|
Mild | R (0, 1a, 1b) | 0 | Endoscopy normal, Imaging Normal | 15 (45.45%) |
1a | Only Nasal mucosa involved, Sinuses clear | |||
1b | Mucosal thickening of One or two sinuses | |||
O (0, 1) | 0 | No orbital involvement, Vision Normal | ||
1 | Mild proptosis/movement restriction, One or Two EOMs enlarged, Vision normal | |||
C (0) | 0 | No neurological signs, Cavernous sinus normal | ||
Moderate | R (2a, 2b) | 2a | One or two sinuses hazy | 06 (18.18%) |
2b | Unilateral pansinusitis with mucosal thickening | |||
O (2a, 2b) | 2a | Moderate proptosis/movement restriction medial/focal extra-conal orbital involvement with contrast enhancement | ||
2b | Moderate proptosis/movement restriction medial/focal extra-conal orbital involvement with no contrast enhancement | |||
C (0) | 0 | No neurological signs, Cavernous sinus normal | ||
Severe | R (3a, 3b) | 3a | Unilateral Sinusitis, completely hazy sinuses | 12 (36.36%) |
3b | Bilateral Sinusitis, complete or incompletely hazy | |||
O (2c, 3a, 3b) | 2c | Localised subperiosteal abscess | ||
3a | Severe proptosis/complete ophthalmoplegia, No vision with diffuse orbital involvement | |||
3b | Severe proptosis/complete ophthalmoplegia, No vision with diffuse orbital involvement with SOV thrombosis | |||
C (1, 2, 3) | 1 | Focal Cavernous sinus involvement | ||
2 | Focal Cavernous sinus involvement with cavernous sinus thrombosis | |||
3 | Unifocal or multifocal CNS disease |
KOH + CFW | Culture | PCR Assay | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
M (+) | M + F (+) | F (+) | M (+) | M + F (+) | F (+) | M (+) | M + F (+) | F (+) | ||
Sample Based Diagnosis (N = 83) | Nasal biopsy (n = 30) | 15 (50.0%) | 02 (6.7%) | 03 (10%) | 07 (23.3%) | 01 (3.3%) | 06 (20.0%) | 03 (10.0%) | 16 (53.3%) | 08 (26.7%) |
Endonasal swabs (n = 27) | 02 (7.4%) | 01 (3.7%) | 01 (3.7%) | 0 (0%) | 0 (0%) | 03 (11.1%) | 03 (11.1%) | 11 (40.7%) | 07 (25.9%) | |
Orbit Biopsy (n = 7) | 02 (28.6%) | 0 (0%) | 0 (0%) | 02 (28.6%) | 0 (0%) | 03 (42.9%) | 02 (28.5%) | 01 (14.3%) | 04 (57.1%) | |
Brain Biopsy (n = 1) | 0 (0%) | 0 (0%) | 0 (0%) | 0 (0%) | 0 (0%) | 0 (0%) | 01 (100%) | 0 (0%) | 0 (0%) | |
Lid Biopsy (n = 1) | 0 (0%) | 01 (100%) | 0 (0%) | 0 (0%) | 01 (100%) | 0 (0%) | 0 (0%) | 01 (100%) | 0 (0%) | |
Control swabs (n= 17) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
Patient Based Diagnosis (N = 33) | 15 (45.5%) | 03 (9.1%) | 04 (12.1%) | 06 (18.2%) | 03 (9.1%) | 07 (21.2%) | 03 (9.1 %) | 17 (51.5%) | 11 (33.3%) |
Sample (No of Patients) | KOH + CFW Positive (%) | KOH + CFW Negative | p Value | PCR Positive | PCR Negative | p Value | |
---|---|---|---|---|---|---|---|
Overall efficacy | Nasal Biopsy (n = 30) | 20 (66.7%) | 10 (33.3%) | NA | 27 (90%) | 03 (10%) | NA |
Endonasal Swab (n = 27) | 04 (14.8%) | 23 (85.2%) | NA | 21 (77.8%) | 06 (22.2%) | NA | |
Comparison of efficacy in select group of patients † | Nasal Biopsy (n = 25) | 17 (68.0%) | 08 (32.0%) | p < 0.001 | 23 (92.0%) | 02 (8.0%) | p = 0.247 |
Endonasal Swab (n = 25) | 03 (12.0%) | 22 (88.0%) | 19 (76.0%) | 06 (24.0%) |
Disease Severity | KOH + CFW Smear (95% CI) | PCR Assay (95% CI) | Tests in Series | Tests in Parallel | |
---|---|---|---|---|---|
Mild | Sn | 100.0% | 100% | 100% | 100% |
Sp | 71.4% (55.3–87.6%) | 42.9% (25.2–60.6%) | 83.2% | 29.8% | |
PPV | 20% (5.7–34.3%) | 11.1% (−0.13–22.4%) | - | - | |
NPV | 100.0% | 100.0% | - | - | |
Moderate to severe | Sn | 75% (60.9–89.1%) | 87.5% (76.7–98.3%) | 65.3% | 100% |
Sp | 78.6% (65.2–92.0%) | 50.0% (33.7–66.3%) | 89% | 39.0% | |
PPV | 50.0% (33.7–66.3%) | 33.3% (17.9–48.7%) | - | - | |
NPV | 91.7% (82.6–100%) | 93.3% (85.2–101.5%) | - | - |
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Mohapatra, S.; Barik, M.R.; Rath, S.; Sharma, S.; Mohapatra, A.; Behera, S.; Acharya, S.; Pattjoshi, D.R.; Padhi, R.K.; Behera, H.S. Diagnostic Performance and Clinical Utility of Conventional PCR Assay in Early Diagnosis of COVID-19 Associated Rhino-Orbito-Cerebral Mucormycosis. J. Fungi 2022, 8, 844. https://doi.org/10.3390/jof8080844
Mohapatra S, Barik MR, Rath S, Sharma S, Mohapatra A, Behera S, Acharya S, Pattjoshi DR, Padhi RK, Behera HS. Diagnostic Performance and Clinical Utility of Conventional PCR Assay in Early Diagnosis of COVID-19 Associated Rhino-Orbito-Cerebral Mucormycosis. Journal of Fungi. 2022; 8(8):844. https://doi.org/10.3390/jof8080844
Chicago/Turabian StyleMohapatra, Samir, Manas Ranjan Barik, Suryasnata Rath, Savitri Sharma, Archisman Mohapatra, Sharmistha Behera, Souvagini Acharya, Dipti Ranjan Pattjoshi, Rajesh Kumar Padhi, and Himansu Sekhar Behera. 2022. "Diagnostic Performance and Clinical Utility of Conventional PCR Assay in Early Diagnosis of COVID-19 Associated Rhino-Orbito-Cerebral Mucormycosis" Journal of Fungi 8, no. 8: 844. https://doi.org/10.3390/jof8080844