Development of a multiplex RT-PCR for simultaneous diagnosis of human metapneumovirus (HMPV) and human respiratory syncytial virus (HRSV) from clinical specimens

• Published in: Virusdisease Vol. 27; no. 4; pp. 375 - 381
• Main Authors: Dayakar, Seetha, Pillai, Heera R., Thulasi, Vineetha P., Nair, Radhakrishnan R.
• Format: Journal Article
• Language: English
• Published: New Delhi Springer India 01.12.2016; Springer Nature B.V
• Subjects: Agreements; Biochemistry; Biomedical and Life Sciences; Cell Biology; Cell culture; Diagnosis; Immunocompromised hosts; Infections; Laboratories; Life Sciences; Manufacturers; Microbiology; Original; Original Article; Pathogens; Patients; Phylogenetics; Polymerase chain reaction; Protein Structure; Real time; Respiratory syncytial virus; Respiratory tract infection; RNA-directed DNA polymerase; Statistical analysis; Viruses; United States > US; Germany; India; Human respiratory syncytial virus; Acute respiratory infection; Multiplex reverse transcriptase PCR; Real-time reverse transcriptase PCR; Human metapneumovirus
• ISSN: 2347-3584; 2347-3517
• DOI: 10.1007/s13337-016-0348-2

Human metapneumovirus (HMPV) and human respiratory syncytial virus (HRSV) are ubiquitous respiratory viral pathogens. They belong to the family Paramyxoviridae (subfamily Pneumovirinae ) and is responsible for acute respiratory tract infections in children, elderly and immunocompromised patients. We designed and tested a multiplex reverse transcriptase polymerase chain reaction (mRT-PCR) as a cost-effective alternative to real-time PCR and cell culture based detection for HMPV and HRSV. The newly developed PCR was used to screen nasal/throat swab samples from 356 patients with suspected acute respiratory infection attending the Government Medical College, Thiruvananthapuram, Kerala, India. The method was compared with a commercially available kit employing real time PCR, for its sensitivity and specificity. 53 (14.9 %) samples were positive for at least one tested pathogen by mRT-PCR. All except one among the positive samples showed similar pathogen profile when tested using real time PCR. 8 (15.1 %) among these 53 were positive for HRSVA, 33 (62.3 %) positive for HRSVB and 12 (22.6 %) were positive for HMPV. 17 (32.7 %) samples showed co-infections in them. Sensitivity and specificity of the mRT-PCR was comparable to that of the commercial kit. Our findings indicate that this newly developed mRT-PCR can be used as a cost-effective alternative for laboratory diagnosis of HMPV/HRSV infection and will significantly reduce diagnostic costs for these viruses in clinical settings.