소아에서 폐렴구균 집락률 측정을 위해 비인두 흡인물의 총 RNA를 이용한 실시간 중합효소 연쇄반응법
Real-time Reverse Transcription Polymerase Chain Reaction Using Total RNA Extracted from Nasopharyngeal Aspirates for Detection of Pneumococcal Carriage in Children
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Purpose: Monitoring pneumococcal carriage rates is important. We developed and evaluated the accuracy of a real-time reverse transcription polymerase chain reaction (RT-PCR) protocol for the detection of Streptococcus pneumoniae . Methods: In October 2014, 157 nasopharyngeal aspirates were collected from patients aged <18 years admitted to Chung-Ang University Hospital. We developed and evaluated a real-time PCR method for detecting S. pneumoniae by comparing culture findings with the results of the real-time PCR using genomic DNA (gDNA). Of 157 samples, 20 specimens were analyzed in order to compare the results of cultures, realtime PCR, and real-time RT-PCR. Results: The concordance rate between culture findings and the results of real-time PCR was 0.922 (P <0.01, Fisher exact test). The 133 culture-negative samples were confirmed to be negative for S. pneumoniae using real-time PCR. Of the remaining 24 culture-positive samples, 21 were identified as S. pneumonia -positive using real-time PCR. The results of real-time RT-PCR and real-time PCR from 20 specimens were consistent with culture findings for all S. pneumoniae -positive samples except one. Culture and real-time RT-PCR required 26.5 and 4.5 hours to perform, respectively. Conclusions: This study established a real-time RT-PCR method for the detection of pneumococcal carriage in the nasopharynx. Real-time RT-PCR is an accurate, convenient, and time-saving method; therefore, it may be useful for collecting epidemiologic data regarding pneumococcal carriage in children.
Abstract
Purpose: Monitoring pneumococcal carriage rates is important. We developed and evaluated the accuracy of a real-time reverse transcription polymerase chain reaction (RT-PCR) protocol for the detection of Streptococcus pneumoniae . Methods: In October 2014, 157 nasopharyngeal aspirates were collected from patients aged <18 years admitted to Chung-Ang University Hospital. We developed and evaluated a real-time PCR method for detecting S. pneumoniae by comparing culture findings with the results of the real-time PCR using genomic DNA (gDNA). Of 157 samples, 20 specimens were analyzed in order to compare the results of cultures, realtime PCR, and real-time RT-PCR. Results: The concordance rate between culture findings and the results of real-time PCR was 0.922 (P <0.01, Fisher exact test). The 133 culture-negative samples were confirmed to be negative for S. pneumoniae using real-time PCR. Of the remaining 24 culture-positive samples, 21 were identified as S. pneumonia -positive using real-time PCR. The results of real-time RT-PCR and real-time PCR from 20 specimens were consistent with culture findings for all S. pneumoniae -positive samples except one. Culture and real-time RT-PCR required 26.5 and 4.5 hours to perform, respectively. Conclusions: This study established a real-time RT-PCR method for the detection of pneumococcal carriage in the nasopharynx. Real-time RT-PCR is an accurate, convenient, and time-saving method; therefore, it may be useful for collecting epidemiologic data regarding pneumococcal carriage in children.
참고문헌 (22)
- Dahlblom V, "Bacterial interactions in the nasopharynx:effects of host factors in children attending daycare centers", J Infect Public Health, 5, pp.133~139 (2012)
- Gray BM, "Epidemiologic studies of Streptococcus pneumoniae in infants: acquisition, carriage, and infection during the first 24 months of life", J Infect Dis 1980;, 142, pp.923~933 (1980)
- Syrjanen RK, "Nasopharyngeal carriage of Streptococcus pneumoniae in Finnish children younger than 2 years old", J Infect Dis, 184, pp.451~459 (2001)
- Levy PY, "Relation between nasal carriage of Staphylococcus aureus and surgical site infection in orthopedic surgery: the role of nasal contamination. A systematic literature review and meta-analysis", Orthop Traumatol Surg Res, 99, pp.645~651 (2013)
- 김선미, "국내 소아의 비강 내 폐구균 보균율과 혈청형에 관한 역학연구", Korean Journal of Pediatrics, 47(6), pp.611~616 (2004), 대한소아과학회
- Paule SM, "Direct detection of Staphylococcus aureus from adult and neonate nasal swab specimens using real-time polymerase chain reaction", J Mol Diagn, 6, pp.191~196 (2004)
- Chan YR, "Molecular diagnostic methods in pneumonia", Curr Opin Infect Dis, 20, pp.157~164 (2007)
- Wood JB, "Nelson textbook of pediatrics", pp.1322~1327 (2016), Elsevier Saunders
- Feigin RD, "Feigin & Cherry's textbook of pediatric infectious diseases", pp.1198~1246 (2013), Saunders
- 신지혜, "소아에서 multiplex RT-PCR에 의한 인후부 상주균 검출", Korean Journal of Pediatrics, 52(12), pp.1358~1363 (2009), 대한소아과학회
- 이미애, "호흡기 검체에서 폐렴의 세균성 원인균 검출을 위한 다중 PCR 검사의 임상적 평가", Annals of Clinical Microbiology, 13(1), pp.40~46 (2010), 대한임상미생물학회
- Al-Marzooq F, "Development of multiplex real-time PCR for the rapid detection of five bacterial causes of community acquired pneumonia", Trop Biomed, 28, pp.545~556 (2011)
- Caliendo AM, "Multiplex PCR and emerging technologies for the detection of respiratory pathogens", Clin Infect Dis, 52(4), pp.S326~S330 (2011)
- Brugger SD, "Detection of Streptococcus pneumoniae strain cocolonization in the nasopharynx", J Clin Microbiol, 47, pp.1750~1756 (2009)
- Carvalho Mda G, "Evaluation and improvement of real-time PCR assays targeting lytA, ply, and psaA genes for detection of pneumococcal DNA", J Clin Microbiol, 45, pp.2460~2466 (2007)
- Simoes AS, "lytA-based identification methods can misidentify Streptococcus pneumoniae", Diagn Microbiol Infect Dis, 85, pp.141~148 (2016)
- Park HK, "Identification of the cpsA gene as a specific marker for the discrimination of Streptococcus pneumoniae from viridans group streptococci", J Med Microbiol, 59(Pt 10), pp.1146~1152 (2010)
- Keith ER, "Characteristics of Streptococcus pseudopneumoniae isolated from purulent sputum samples", J Clin Microbiol, 44, pp.923~927 (2006)
- Kurola P, "Presence of capsular locus genes in immunochemically identified encapsulated and unencapsulated Streptococcus pneumoniae sputum isolates obtained from elderly patients with acute lower respiratory tract infection", J Med Microbiol, 59(Pt 10), pp.1140~1145 (2010)
- Stensballe LG, "Comparison of nasopharyngeal aspirate and nasal swab specimens for detection of respiratory syncytial virus in different settings in a developing country", Trop Med Int Health, 7, pp.317~321 (2002)
- Ohrmalm L, "Flocked nasal swab versus nasopharyngeal aspirate for detection of respiratory tract viruses in immunocompromised adults: a matched comparative study", BMC Infect Dis, 10, pp.340~ (2010)
- Satzke C, "Standard method for detecting upper respiratory carriage of Streptococcus pneumoniae:updated recommendations from the World Health Organization Pneumococcal Carriage Working Group", Vaccine, 32, pp.165~179 (2013)