Sains Malaysiana 47(11)(2018): 2821–2829

http://dx.doi.org/10.17576/jsm-2018-4711-25

 

Multiplex Real-Time PCR Detection of Respiratory Viruses in Lower Respiratory Tract Infections in Children

(Multipleks PCR Masa Sebenar untuk Pengesanan Jangkitan Virus Pernafasan dalam Saluran Pernafasan Bahagian Bawah bagi Kanak-Kanak)

 

ZETTI ZAINOL RASHID^1*, PEI CHUEN LEE², UMI KALSOM ALI¹, MUTTAQILLAH NAJIHAN ABDUL SAMAT¹ & SWEE FONG TANG²

 

¹Department of Medical Microbiology & Immunology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, 56000 Kuala Lumpur, Federal Territory, Malaysia

 

²Department of Paediatrics, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, 56000 Kuala Lumpur, Federal Territory, Malaysia

 

Received: 24 May 2018/Accepted: 3 July 2018

 

ABSTRACT

Acute respiratory infections cause significant morbidity and mortality in children. Several new respiratory viruses have been identified and co-detection of multiple viruses is commonly reported. This is part of a prospective study which aims to detect respiratory viruses by multiplex molecular method and conventional methods. Nasopharyngeal aspirate specimens were taken from hospitalised children aged less than 5 years with lower respiratory tract infections. These were tested using viral culture, immunofluorescence and Seegene Anyplex™ II RV16 real-time polymerase chain reaction. From 102 samples, 69 (67.6%) were positive by PCR, 12 (11.8%) positive by culture method and 13 (12.7%) positive by IF. A single viral pathogen was detected in 48 samples (47.1%), while 21 samples (20.6%) had co-detection of 2 to 4 viral pathogens. Respiratory syncytial virus (RSV) was most common, detected in 17 samples (16.7% of all samples), followed by adenovirus and rhinovirus in 16 (15.7%), respectively. Bocavirus was detected in 15, enterovirus in 15, influenza A in 8 and parainfluenza-4 in 4 samples, with highest occurrences in co-detection (12/15, 10/15, 5/8 and 3/4, respectively). RSV was the least likely detected in co-detection (3/17). In PCR-positive samples, 54/69 (78.3%) were patients aged up to 24 months. Molecular methods detect more viral aetiologies than conventional methods, with simultaneous detection of multiple respiratory viruses. More sensitive, specific and rapid tools to determine aetiological agents could be incorporated into diagnostic algorithms of respiratory tract infections. Interpretations, significance, and applicability in clinical practice could be further explored, particularly for patients up to 2 years old.

 

Keywords: Infections; molecular; multiplex; PCR; respiratory viruses

 

ABSTRAK

Jangkitan pernafasan akut mengakibatkan morbiditi dan mortaliti yang ketara pada kanak-kanak. Beberapa virus pernafasan yang baru telah dikenal pasti dan pengesanan beberapa virus secara serentak biasa dilaporkan. Kajian ini adalah sebahagian daripada kajian prospektif bertujuan untuk mengesan virus pernafasan menggunakan kaedah molekul multipleks dan kaedah konvensional. Spesimen aspirat nasofarinks diambil daripada kanak-kanak berumur kurang daripada 5 tahun yang dimasukkan ke hospital kerana jangkitan saluran pernafasan bahagian bawah. Pengujian adalah menggunakan pengkulturan virus, imunofluoresen dan reaksi rantai polimerase masa sebenar menggunakan Seegene Anyplex ™ II RV16. Daripada 102 sampel, 69 (67.6%) positif PCR, 12 (11.8%) positif dengan kaedah kultur virus dan 13 (12.7%) positif dengan imunofluoresen. Patogen virus tunggal dikesan dalam 48 sampel (47.1%), manakala 21 sampel (20.6%) mempunyai pengesanan bersama 2 hingga 4 patogen virus. Virus pernafasan sinsitium (RSV) paling banyak dikesan, iaitu dalam 17 sampel (16.7% daripada semua sampel), diikuti oleh adenovirus dan rhinovirus dalam 16 (15.7%) masing-masing. Bocavirus dikesan dalam 15, enterovirus dalam 15, influenza A dalam 8 dan parainfluenza-4 dalam 4 sampel dengan kes tertinggi dalam pengesanan bersama (12/15, 10/15, 5/8 dan 3/4 masing-masing). RSV paling sedikit dikesan dalam pengesanan bersama (3/17). Dalam sampel positif PCR, 54/69 (78.3%) adalah pesakit berusia sehingga 24 bulan. Kaedah molekul mengesan lebih banyak virus daripada kaedah konvensional dengan pengesanan serentak pelbagai virus pernafasan. Kaedah yang lebih sensitif, khusus dan cepat untuk menentukan agen penyebab boleh dimasukkan ke dalam algoritma diagnosis jangkitan saluran pernafasan. Tafsiran keputusan ujian dan kegunaannya dalam amalan klinikal boleh diterokai dengan lebih lanjut, terutama untuk pesakit berusia sehingga 2 tahun.

 

Kata kunci: Jangkitan; molekul; multipleks; PCR; virus pernafasan

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*Corresponding author; email: zetti@ppukm.ukm.edu.my