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. 2018 Winter;13(1):54-62.

Development of Multiplex Reverse Transcription-Polymerase Chain Reaction for Simultaneous Detection of Influenza A, B and Adenoviruses

Mohsen Nakhaie  1 , Hoorieh Soleimanjahi  1 , Hamid Reza Mollaie  2 , Seyed Mohamad Ali Arabzadeh  2
Affiliations

Development of Multiplex Reverse Transcription-Polymerase Chain Reaction for Simultaneous Detection of Influenza A, B and Adenoviruses

Mohsen Nakhaie et al. Iran J Pathol. 2018 Winter.

Abstract

Background and objective: Millions of people in developing countries lose their lives due to acute respiratory infections, such as Influenza A & B and Adeno viruses. Given the importance of rapid identification of the virus, in this study the researchers attempted to design a method that enables detection of influenza A, B, and adenoviruses, quickly and simultaneously. The Multiplex RT PCR method was the preferred method for the detection of influenza A, B, and adenoviruses in clinical specimens because it is rapid, sensitive, specific, and more cost-effective than alternative methods.

Methods: After collecting samples from patients with respiratory disease, virus genome was extracted, then Monoplex PCR was used on positive samples and Multiplex RT-PCR on clinical specimens. Finally, by comparing the bands of these samples, the type of virus in the clinical samples was determined.

Results: Performing Multiplex RT-PCR on 50 samples of respiratory tract led to following results; flu A: 12.5%, fluB: 50%, adeno: 27.5%, negative: 7.5%, and 2.5% contamination.

Conclusion: Reverse transcription-multiplex Polymerase Chain Reaction (PCR) technique, a rapid diagnostic tool, has potential for high-throughput testing. This method has a significant advantage, which provides simultaneous amplification of numerous viruses in a single reaction. This study concentrates on multiplex molecular technologies and their clinical application for the detection and quantification of respiratory pathogens. The improvement in diagnostic testing for viral respiratory pathogens effects patient management, and leads to more cost-effective delivery of care. It limits unnecessary antibiotic use and improves clinical management by use of suitable treatment.

Keywords: Adenovirus; Influenza A; Influenza B; Multiplex RT-PCR; Polymerase chain reaction; Respiratory infection.

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Figures

Figure 1
Figure 1
Positive Sample Bands for Flu A, B and Adenovirus
Figure 2.
Figure 2.
Serial Ten-Fold Dilutions of the Influenza Type A RNA Prepared in Elution Buffer
Figure 3
Figure 3
Typing and Subtyping of Influenza and Adeno Virus in a Panel of Clinical Specimens by Multiplex Real Time-Polymerase Chain Reaction
See this image and copyright information in PMC

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