Abdullah YAZAR, Ebru AYPAR, Recep KEŞLİ, Fatih AKIN, Melike KESER, Dursun ODABAŞ, Muhammet Güzel KURTOĞLU, Ahmet SERT, Hüseyin BİLGİN

Konya bölgesinde toplum kökenli pnömoni için hastaneye yatırılan çocuk hastalarda solunumsal viral enfeksiyon etkenlerinin multipleks gerçek zamanlı-PCR ile tanımlanması

AMAÇ: Bu çalışmanın amacı; toplum kökenli pnömoni tanısı ile hastaneye yatırılan çocuklarda viral pnömoni etkenlerinin moleküler tanı teknikleri kullanılarak tanımlanmasını değerlendirmektir. YÖNTEMLER: Çalışmaya, Konya bölgesindeki viral pnömoni etkenlerini tanımlamak amacı ile toplum kökenli pnömoni nedeni ile hastaneye yatırlan 64 çocuk dahil edilmiştir. Çalışma, Ekim 2009 ve Aralık 2010 tarihleri arasında gerçekleştirilmiştir. Yaşları bir ay ile sekiz yaş arasında olan ve toplum kökenli pnömoni olarak tanı konulan çocuk hastalardan nazo-faringeal sürüntü örnekleri alınmıştır. Hastalara hastaneye yatışları sırasında respiratuvar sinsityal virüs (RSV), adenovirus, influenza virüs ve paranifluenza virüsler gibi viral pnömoni etkenlerinin varlıklarını tepit etmek için multipleks gerçek zamanlı polimeraz zincir reaksiyonu (PZR) testi yapılmıştır. BULGULAR: Toplam 64 hastanın 22 (%34,4)si viral enfeksiyonlar, beş (%7,7)si bakteriyel enfeksiyonlar ve iki (%3,1)i hem viral hem de bakteriyel enfeksiyonlar için pozitif olarak bulunmuştur. 20 (%31,2) vakada sadece viral patojen ve üç (%4,6) hastada sadece bakteriyel enfeksiyon belirlenmiştir. RSV (%23,4) ve adenovirüs (%4,6) en sık viral patojenler olarak tespit edilirken, influenza A virüsü (%3,1), parainfluenza tip 1, 2, 3 virüsleri (%3,1) enfeksiyon nedeni diğer virüsler olarak belirlenmiştir. İnfluenza B virüsü vakaların hiçbirinde tespit edilmemiştir. 37 (%57,9) hastada muhtemel hiçbir etken belirlenememiştir. Çalışma süresince ölüm olmamıştır. SONUÇ: Bu çalışmada, hastaneye yatırılan bir ay ve sekiz yaş arasında çocuklarda toplum kökenli pnömoninin en sık etkeninin virüsler olduğu belirlenmiştir. Virüslerin etiyolojik tanılarının geliştirilmesi, tedavilerde özellikle gereksiz antibiyotik kullanılması ve gereksiz izolasyon uygulamaları yapılmasını önlemede yararlı olacaktır. Hastaneye yatırılan çocuk hastalarda toplum kökenli pnömoninin etiyolojisinde virüslerin rollerini açıklamaya odaklanan daha fazla ve yeni çalışmaların gerçekleştirilmesi bu hasta grubunun tedavisinde doğru politikaların belirlenmesine katkı sağlayacaktır.

Identification of respiratory viral infection agents by multiplex real-time PCR among children hospitalized for community-acquired pneumonia in Konya province

OBJECTIVE: Aim of this study was to evaluate the identification of viral pneumonia agents among children who were diagnosed and hospitalized for community-acquired pneumonia (CAP) by using molecular diagnostic techniques. METHODS: Sixty four children hospitalized for community-acquired pneumonia were included in the study in order to identify viral pneumonia agents in the province of Konya. The study was carried out in between October 2009 and December 2010. Nasopharyngeal smears were obtained from children patients aged 1 month to 8 years who were diagnosed as community-acquired pneumonia. Multiplex real time-Polymerase Chain Reaction (PCR) was performed for detection of existence of viral pneumonia agents such as respiratory syncytial virus (RSV), adenovirus, influenza and parainfluenza viruses on admission. RESULTS: Of all the 64 patients, 22 (34.4%) were positive for viral infections, 5 (7.7 %) were positive for bacterial infections, and 2 (3.1%) were positive for both viral and bacterial infections. A sole viral infection was identified in 20 (31.2%) and a sole bacterial infection in 3 (4.6%) cases. Influenza A virus (3.1%), parainfluenza type 1, 2, 3 viruses (3.1%), respiratory syncytial virus (23.4%) and adenovirus (4.6%) were found as causative viruses. Influenza B virus was not detected in any of cases. No possible etiologic agent was found in 37 cases (57.9%). Respiratory syncytial virus (23.4%) and adenovirus (4.6%) were the most commonly detected viral pathogens. There was no mortality during the study. CONCLUSION: This study showed that the viruses commonly detected as the causative agents of community-acquired pneumonia among the hospitalized children aged between 1 month and 8 years. Improving the etiological diagnosis of viral infections may definitely contribute to avoid unnecessary therapy, particularly antibiotics and allow for preventive isolation of infected patients. Performing more and new studies focusing on defining the role of viruses at the etiology of community-acquired pneumonia among hospitalized children will help establishment of true policies on the treatment of these patients.

PDF

___

1. Wardlaw T, Salama P, Johansson EW, Mason E. Pneumonia: the leading killer of children. Lancet, 2006; 368 (9541): 1048-50.
2. Glezen WP, Loda FA, Clyde WA Jr, Senior RJ, Sheaffer CI, Conley WG, et al. Epidemiologic patterns of acute lower respiratory disease of children in a pediatric group practice. J Pediatr, 1971; 78 (3): 397- 406.
3.Henrickson KJ. Viral pneumonia in children. Semin Pediatr Infect Dis, 1998; 9: 217-33.
4. Sinaniotis CA, Sinaniotis AC. Community-acquired pneumonia in children. Curr Opin Pulm Med, 2005; 11 (3): 218-25.
5. Aguilar JC, Pérez-Breña MP, García ML, Cruz N, Erdman DD, Echevarría JE. Detection and identification of human parainfluenza viruses 1, 2, 3, and 4 in clinical samples of pediatric patients by multiplex reverse transcription-PCR. J Clin Microbiol, 2000; 38 (7): 1191-5.
6. Coiras MT, Pérez-Breña P, García ML, Casas I. Simultaneous detection of influenza A, B, and C viruses, respiratory syncytial virus, and adenoviruses in clinical samples by multiplex reverse transcription nested-PCR assay. J Med Virol, 2003; 69 (1): 132-44.
7. Fan J, Henrickson KJ, Savatski LL. Rapid simultaneous diagnosis of infections with respiratory syncytial viruses A and B, influenza viruses A and B, and human parainfluenza virus types 1, 2, and 3 by multiplex quantitative reverse transcription-polymerase chain reaction-enzyme hybridization assay (Hexaplex). Clin Infect Dis, 1998; 26 (6): 1397-402.
8. Gröndahl B, Puppe W, Hoppe A, Kühne I, Weigl JA, Schmitt HJ. Rapid identification of nine microorganisms causing acute respiratory tract infections by single- tube multiplex reverse transcription-PCR: feasibility study. J Clin Microbiol, 1999; 37 (1): 1-7.
9. Liolios L, Jenney A, Spelman D, Kotsimbos T, Catton M, Wesselingh S. Comparison of a multiplex reverse transcription-PCR-enzyme hybridization assay with conventional viral culture and immunofluorescence techniques for the detection of seven viral respiratory pathogens. J Clin Microbiol, 2001; 39 (8): 2779-83.
10.Templeton KE, Scheltinga SA, Beersma MF, Kroes AC, Claas EC. Rapid and sensitive method using multiplex real-time PCR for diagnosis of infections by influenza A and influenza B viruses, respiratory syncytial virus, and parainfluenza viruses 1, 2, 3, and 4. J Clin Microbiol, 2004; 42 (4): 1564-9.
11. McIntosh K. Community-acquired pneumonia in children. N Engl J Med, 2002; 346 (6): 429-37.
12.British Thoracic Society. Guidelines for the management of community-acquired pneumonia in childhood. Thorax, 2002; 57 (Suppl 1): 1-24.
13. World Health Organization. Technical bases for the WHO recommendation on the management of pneumonia in children at first level health facilities, 1991; Geneva.
14.Juvén T, Mertsola J, Waris M, Leinonen M, Meurman O, Roivainen M, et al. Etiology of community- acquired pneumonia in 254 hospitalized children. Pediatr Infect Dis J, 2000; 19 (4): 2938.
15.Turner RB, Lande AE, Chase P, Hilton N, Weinberg D. Pneumonia in pediatric outpatients: cause and clinical manifestations. J Pediatr, 1987; 111 (2): 194-200.
Nohynek H, Eskola J, Laine E, Halonen P, Ruutu P, Saikku P, et al. The causes of hospital treated lower respiratory tract infection in children. Am J Dis Child, 1991; 145 (6): 618-22.
17.Ruuskanen O, Nohynek H, Ziegler T, Capeding R, Rikalainen H, Huovinen P, et al. Pneumonia in childhood: etiology and response to antimicrobial therapy. Eur J Clin Microbiol Infect Dis, 1992; 11 (3): 217-23.
18.Heiskanen-Kosma T, Korppi M, Jokinen C, Kurki S, Heiskanen L, Juvonen H, et al. Etiology of childhood pneumonia: serologic results of a prospective, population-based study. Pediatr Infect Dis J, 1998; 17 (11): 986-91.
19. Wubbel L, Muniz L, Ahmed A, Trujillo M, Carubelli C, McCoig C, et al. Etiology and treatment of community- acquired pneumonia in ambulatory children. Pediatr Infect Dis J, 1999; 18 (2): 98-104.
20.Claesson BA, Trollfors B, Brolin I, Granström M, Henrichsen J, Jodal U, et al. Etiology of community acquired pneumonia in children based on antibody responses to bacterial and viral antigens. Pediatr Infect Dis J, 1989; 8 (12): 856-62.
21. Cilla G, Oñate E, Perez-Yarza EG, Montes M, Vicente D, Perez-Trallero E. Viruses in community- acquired pneumonia in children aged less than 3 years old: High rate of viral coinfection. J Med Virol, 2008; 80 (10): 1843-9.
22.Henrickson KJ, Hoover S, Kehl KS, Hua W. National disease burden of respiratory viruses detected in children by polymerase chain reaction. Pediatr Infect Dis J, 2004; 23 (Suppl 1): 11-8.
23. Samransamruajkit R, Hiranrat T, Chieochansin T, Sritippayawan S, Deerojanawong J, Prapphal N, et al. Prevalence, clinical presentations and complications among hospitalized children with influenza pneumonia. Jpn J Infect Dis, 2008; 61 (6): 446-9.
24. Iwane MK, Edwards KM, Szilagyi PG, Walker FJ, Griffin MR, Weinberg GA, et al. Population-based surveillance for hospitalizations associated with respiratory syncytial virus, influenza virus, and parainfluenza viruses among young children. Pediatrics, 2004; 113 (6): 1758-64.
25. Michelow IC, Olsen K, Lozano J, Rollins N. Epidemiology and characteristics of community acquired pneumonia in hospitalized children. Pediatrics, 2004; 113 (4): 701-7.