Viral pnömoni tanısı alan hastalarda etkenlerin multipleks PZR yöntemiyle gösterilmesi

Abstract

Amaç: Çalışmanın amacı Selçuk Üniversitesi Meram Tıp Fakültesi Hastanesi'ne pnömoni nedeniyle başvuran çocuk ve yetişkin hastalarda, viral etkenlerin multiplex polimeraz zincir reaksiyonuyla tespit edilmesidir.Gereç ve Yöntem: Ekim 2010 ? Nisan 2011 tarihleri arasında hastaneye başvuran150 çocuk ve 150 yetişkin pnömonili hastadan nazofarengeal sürüntü örneği toplandı. Bu örnekler çalışılıncaya kadar -20°C'de saklandı. Viral DNA/RNA ekstraksiyonu ve amplifikasyonu Seeplex®RV12 multiplex PZR kiti kullanılarak yapıldı. Viral DNA/ RNA çoğaltıldıktan sonra etidyum bromid ile boyanıp %2 agaroz jelde yürütüldü ve büyüklüklerine göre viruslar identifiye edildi.Bulgular: Hastaların 150(%50)'si çocuk kliniğinden, 150(%50)'si ise Göğüs Hastalıkları Kliniği'nden gönderildi. Tüm hastaların %46.2'sinde, en az 1 virus tespit edildi. Pozitif hastaların %45'i RSV olarak tespit edildi. Bunu %19.4 ile rinovirüs, %13.6 ile influenza A, %6.5 ile parainfluenza, %4.3 ile influenza B, %3.6 ile adenovirüs, %2.9 ile koronavirüs ve %1.4 ile metapnömovirus izledi. Toplam 139 olan pozitifin 107'si çocuk ve 32'si yetişkindi. 5 (%1.6) hastada ko-infeksiyon izlendi. 0-4 yaş grubunun %71'inde ve 5-9 yaş grubunun %75'inde etken virus bulundu. Erişkinde virus bulunma oranı düşüktü. Bu farklılık istatistiksel olarak anlamlı bulundu(p> 0.537).Sonuç: Bizim çalışmamızda %44.6 RSV, %19.4 rinovirus, %17.9 influenza virus, %6.5 parainfluenza virus, %3.6 adenovirus, %2.9 koronavirus, %1.4 metapnömovirus tespit edildi.Multipleks PZR testleri, solunum yolu infeksiyonları için kolay, hızlı ve güvenli testlerdir. Bu testlerin, klinik mikrobiyoloji laboratuvarlarında rutin tanı testleri arasında yer alması gereklidir. Viral hastalıklarda etkenin tespit edilmesi, gereksiz antibiyotik reçetelenmesinin önlenmesi ve gerektiğinde uygun antiviralin verilmesini sağlar. Anahtar kelimeler: Solunum yolu infeksiyonları, multipleks PZR, nazofarengeal sürüntü.

Objective: The aims of this study was to determine the viral etiology of pneumonia among children and adults at Selcuk University Meram Medical Faculty Hospital using multiplex Polymerase Chain Reaction (PCR) assay.Materials And Methods: Between October 2010-April 2011, nasopharyngeal swab samples were collected from 150 children and 150 adults with pneumonia. All the samples were stored in a freezer at -20°C until PCR was carried out. Viral DNA/RNA was extracted using Viral Gene Spin? kit according to the manufacturer?s instructions and was amplified by Seeplex®RV12 multiplex PCR kit. The amplified products were visualized by electrophoresis an ethidium bromide-stained 2% agarose jel and analyzed by identifying bands.Results: 150 patients were from pediatric clinics and 150 patients were from pulmonary diseases clinics. At least one viral agent was detected in 46.2% of all patients. RSV was the most common agents encountered among virus DNA positive patients (45%), and followed by rhinovirus (19.4%), influenza A (13.6%), parainfluenza (6.5%), influenza B(4.3%), adenovirus (3.6%), koronavirus (2.9%) and human metapneumovirus (1.4%) viruses. Positive in 107 (35.6%) of the 139 patients were children and 32 (10.6%) were adults. Co-infection was detected in 5 (1.6%) of all patients. The rate of virus positive was found to be 71% in age group of 0-4 and was found to be 75% in age group of 5-9. The rate of virus positive in adults was lower. This difference was found statistically significant (p> 0.537).Conclusion: In this study, the rate of RSV was found to be 45%, rinovirus was found to be 19.4%, influenza virus was found to be 18%, parainfluenza was found to be 6.5%, adenovirüs was found to be 3.6%, koronavirüs was found to be 2.9 % and metapnömovirus was found to be 1.4%.Multiplex PCR assays are rapid, easy and reliable methods and used in the diagnosis of respiratory diseases. These tests are requared to take place among the routine diagnostic procedurs at clinical microbiology laboratories. Viral detection may reduce unnecessary prescription of antibiotics and may prompt antiviral treatment such as oseltamivir (against influenza) to the patient or exposed family members.