Yayın: Solunum yolu örneklerinden saptanan respiratuvar sinsityal virüslerin moleküler yöntemlerle tiplendirilmesi
Dosyalar
Tarih
Kurum Yazarları
Yazarlar
Yağdı, Oğuzhan
Danışman
Ağca, Harun
Dil
Türü
Yayıncı:
Bursa Uludağ Üniversitesi
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Özet
Amaç: Nazofaringeal sürüntü örneklerinde multipleks polimeraz zincir reaksiyonuyla (PZR) tespit edilen Respiratuvar sinsityal virüslerin (RSV) moleküler yöntemlerle tiplendirilmesi, tiplerin mevsimsel dağılım, yaş, cinsiyet kategorileriyle ilişkisinin belirlenmesi, elde edilen verilerin epidemiyolojiye katkı sağlamasıdır. Gereç ve Yöntem: Multipleks PZR yöntemiyle RSV varlığı tespit edilmiş, -80°C’de saklanan, 0-90 yaş hastalara ait nazofaringeal sürüntü örneklerinden viral RNA izolasyonu sonrası cDNA sentezi gerçekleştirildi. Hedeflenen bölge PZR ile çoğaltılarak jel elektroforezi ile doğrulandı ve pürifikasyon işlemi uygulandı. Sekanslama işlemi, Sanger dideoksi dizileme yöntemiyle gerçekleştirildi ve elde edilen diziler biyoinformatik araçlarla analiz edildi. Alt tip belirleme, filogenetik ağaç oluşturma, dağılım analizleri yapılarak veriler istatistiksel olarak değerlendirildi. Bulgular: Tiplendirilebilen 52 örneğin %71’i RSV-A, %29’u RSV-B olarak belirlendi. RSV-A alt grubunun tamamı GA.2.3.5, RSV-B alt grubunun tamamı GB.5.0.5a genotipine ait olup en sık rastlanan soylar sırasıyla RSV-A için A.D.1 (%78) ve RSV-B için B.D.E.1 (%80) olarak tespit edildi. Aralık ayında RSV-A saptanma ihtimali, ocak ayında ise RSV-B saptanma ihtimali istatistiksel anlamlı bulundu (p=0,0138, p=0,0024). 2025 yılında RSV-B alt grubunda artış istatistiksel olarak anlamlı bulundu (p=0,0001). Sonuç: Çalışmamız, bölgemizde RSV-A GA.2.3.5 ve RSV-B GB.5.0.5a genotiplerinin baskın olduğunu gösterdi. Pandemi sonrasında, küresel artış eğilimiyle uyumlu şekilde RSV-B’nin artışı ve RSV-B için hakim soyun B.D.E.1 olduğu gösterildi. Çalışmamız, RSV için 2024’te önerilen standart filogenetik sınıflamanın ve RSV aşılarının kullanım onayı almasının ardından ülkemizde yapılan ilk RSV tiplendirme çalışmasıdır. Çalışma, RSV epidemiyolojisi ve dinamikleri hakkında önemli veriler sağlarken, aşıların kullanıma girmesi halinde elimizde temel verilerin olmasına katkı sağlayarak, yeni aşılama ve enfeksiyon kontrol stratejileri için bilimsel bir temel oluşturmaktadır.
Objective: This study aims to type Respiratory Syncytial Viruses (RSV) detected in nasopharyngeal swab samples using multiplex polymerase chain reaction (PCR), analyze their seasonal distribution, and investigate their association with age and gender categories by molecular methods and contribute to epidemiology of RSV. Materials and Methods: Viral RNA was isolated from nasopharyngeal swab samples collected from patients aged 0-90 years, which had been stored at - 80°C and detected positive for RSV using the multiplex PCR method. cDNA synthesis was performed, and the target region was amplified by PCR, validated through gel electrophoresis, and purified. Sequencing was conducted using the Sanger dideoxy method, and the sequences were analyzed with bioinformatics tools. Subtype identification, phylogenetic tree construction, and distribution analyses were performed, and the data were statistically analysed. Results: Of the 52 typable samples, 71% were identified as RSV-A and 29% as RSV-B. All RSV-A strains belonged to the GA.2.3.5 genotype, while all RSV-B strains belonged to the GB.5.0.5a genotype. The most common lineages were A.D.1 (78%) for RSV-A and B.D.E.1 (80%) for RSV-B. The detection probability of RSV-A in December and RSV-B in January was statistically significant (p=0,0138, p=0,0024). In 2025, a statistically significant increase was observed in the prevalence of RSV-B (p=0,0001). Conclusion: Our study demonstrated that RSV-A GA.2.3.5 and RSV-B GB.5.0.5a genotypes are predominant in our region. After the pandemic, an increase in RSV-B cases, consistent with the global upward trend, was observed, with B.D.E.1 identified as the dominant lineage for the RSV-B subgroup. This study represents the first RSV typing research conducted in our country following the implementation of the standardized phylogenetic classification proposed in 2024 and the approval of RSV vaccines. While providing essential data on RSV epidemiology and dynamics, this study contributes to the availability of baseline data in case of vaccine implementation and establishes a scientific basis for new vaccination and infection control strategies.
Objective: This study aims to type Respiratory Syncytial Viruses (RSV) detected in nasopharyngeal swab samples using multiplex polymerase chain reaction (PCR), analyze their seasonal distribution, and investigate their association with age and gender categories by molecular methods and contribute to epidemiology of RSV. Materials and Methods: Viral RNA was isolated from nasopharyngeal swab samples collected from patients aged 0-90 years, which had been stored at - 80°C and detected positive for RSV using the multiplex PCR method. cDNA synthesis was performed, and the target region was amplified by PCR, validated through gel electrophoresis, and purified. Sequencing was conducted using the Sanger dideoxy method, and the sequences were analyzed with bioinformatics tools. Subtype identification, phylogenetic tree construction, and distribution analyses were performed, and the data were statistically analysed. Results: Of the 52 typable samples, 71% were identified as RSV-A and 29% as RSV-B. All RSV-A strains belonged to the GA.2.3.5 genotype, while all RSV-B strains belonged to the GB.5.0.5a genotype. The most common lineages were A.D.1 (78%) for RSV-A and B.D.E.1 (80%) for RSV-B. The detection probability of RSV-A in December and RSV-B in January was statistically significant (p=0,0138, p=0,0024). In 2025, a statistically significant increase was observed in the prevalence of RSV-B (p=0,0001). Conclusion: Our study demonstrated that RSV-A GA.2.3.5 and RSV-B GB.5.0.5a genotypes are predominant in our region. After the pandemic, an increase in RSV-B cases, consistent with the global upward trend, was observed, with B.D.E.1 identified as the dominant lineage for the RSV-B subgroup. This study represents the first RSV typing research conducted in our country following the implementation of the standardized phylogenetic classification proposed in 2024 and the approval of RSV vaccines. While providing essential data on RSV epidemiology and dynamics, this study contributes to the availability of baseline data in case of vaccine implementation and establishes a scientific basis for new vaccination and infection control strategies.