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Short Commentry | DOI: https://doi.org/10.31579/2690-8808/115
1 Professor of Pathology, Consultant Microbiologist, Al Nafees Medical College & Hospital, Islamabad, Pakistan.
2 Professor of Pathology, Consultant Microbiologist, National University of Medical Sciences, Rawalpindi. Pakistan.
3 Assistant Professor of Pathology, Consultant Microbiologist, Armed Forces Institute of Cardiology & National Institute of Heart Diseases, National University of Medical Sciences, Rawalpindi. Pakistan.
4 Associate Professor of Pathology, Consultant Microbiologist, Armed Forces Institute of Pathology, National University of Medical Sciences,Rawalpindi. Pakistan
*Corresponding Author: Humaira Zafar, Professor of Pathology, Consultant Microbiologist, Al Nafees Medical College & Hospital, Islamabad, Pakistan.
Citation: Humaira Zafar, Irfan Ali Mirza, Muhammad Fayyaz and Wajid Hussain (2022). Multiplex PCR via Microarray a Surrogate For Rapid Molecular Detection of Mycobacterial and Fungal Species Along with Ant Microbial Resistant Genes, Directly From Clinical Specimens’ Journal of Clinical Case Reports and Studies 3(5); DOI: 10.31579/2690-8808/115
Copyright: © 2022 Humaira Zafar, This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Received: 09 April 2022 | Accepted: 25 April 2022 | Published: 04 June 2022
Keywords: multi drug resistant; mycobacterial; invasive fungal infections
Multi drug resistant Mycobacterial and invasive fungal infections imposes immense Global health burden resulting in high morbidity and mortality rates. The biggest delima is the time consuming culture and sensitivity diagnostics i.e 04 to 06 weeks for Mycobacterial and 02 to 04 weeks for fungal infections. The delay in result provision adds up to patient’s miseries especially for resistant cases. Therefore the aim of writing this short commentary was to introduce multiplex PCR via microarray bead hybridization assay. A thorough literature review was found extremely deficient. However, a limited published data concluded this technique as a surrogate for rapid molecular and resistant gene depiction for mycobacterial and fungal species directly from clinical specimens. Another biggest advantage was simultaneous genomics and resistant genes identification for Mycobacterial and Fungal isolates. The resistant genes for 1st line anti tuberculous therapy (ATT), 2nd line ATT and commonly used anti fungals can be detected easily. This cost effective test can be finalized in approximately less than 24 hours. Thus early and accurate management can be timely started for both said infections.
The Global health care providers are facing a biggest challenge to manage Multidrug-resistant tuberculosis (MDR-Tb) infections. The situation further worsens, if underlying immunodeficiency becomes a major contributing factor for said infections. Here comes the significance to exploit new, rapid genomic diagnostic and therapeutic modalities to accurately diagnose such cases. This approach comes in lime light as promising lifesaving one [1,2].Currently available gold standard to identify the microbial species and drug susceptibility includes culture and sensitivity. Which unfortunately requires a minimum time period of 04 to 06 weeks for Mycobacterial infections. In view to get early diagnosis, Aslam B from Pakistan in 2017, published a review article emphasizing the utility of microarray for rapid microbial genomic and proteomic analysis. It was discovered that Microarray chips functions for large-scale analysis of whole transcriptome, along with identification of simultaneous genomic and resistant genes of drugs. A high sensitivity throughput and rapid expression analysis are the added benefits, which ranks it superior to western blot and ELISA studies [3].
A published review article by Pérez MA etal in 2022, highlighted the significance for hybridization based microarray in microbiology. He disclosed that after DNA extraction for Mycobacterial species, further probe analysis can be done using a well-designed hybridization microarray i.e multiplex PCR. It has proven to be less complex, cost effective. Furthermore multiple genes can easily be detected in one well of microtiter plate [4]. In 2019, Chen CY carried out the original study in China on pure isolates of Mycobacterium tuberculosis by microarray identification. He designed 12 pairs of primers and 60 single nucleotide polymorphisms of 9 genes i.e isoniazid (inhA), rifampicin (rpoB), pyrazinamide (pncA), ethambutol (embB), kanamycin (katG), streptomycin (rpsL & rrs), flouroquinolones (gyrA & gyrB), and amikacin (eis). The successful results had given a ray of hope for early identification of resistant genes of ATT [5]. Another study concluded the utility of microarray for rapid genomic detection of fungal isolates. The mutation of TR34/L98H in CYP51A gene for Aspergillus fumigatus during posaconazole prophylaxis was identified. ERG11 and ERG3 gene polymorphisms and susceptibility to fluconazole (Azoles) in Candida isolates [6]. Point mutations in Candida glabrata FUR1 (CgFUR1) and CgFKS2, as well as overexpression of CgCDR1 and CgCDR2, linked with fluconazole and voriconazole (Azoles) resistance [6,7].
So far there is extremely limited data regarding utility of multiplex PCR via microarray for genomic (species along with resistant genes) and proteiomic identification of Mycobacterial and fungal isolates. Even the available literature has focused working on pure culture growth of Mycobacterium tuberculosis and few fungi. While non tuberculous Mycobacterium and many fungal species were not studied. The current short commentary has thrown light for the utilization of this under utilized technique in microbiology. Knowing the details for species along with resistant genes for 1st line anti tuberculous therapy (ATT), 2nd line ATT and commonly used anti fungals, in less than 24hrs time will be helpful to reduce the morbidity and mortality rates from said infection
A multiplex PCR via microarray is a surrogate for rapid molecular and resistant gene depiction for mycobacterial and fungal species directly from clinical specimens. This under utilized technique harbours great utility in field of molecular genetics and microbiology.
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