Real-time Reverse Transcription Polymerase Chain Reaction Using Total RNA Extracted from Nasopharyngeal Aspirates for Detection of Pneumococcal Carriage in Children
Pediatric Infection & Vaccine
;
: 194-201, 2016.
Article
in Korean
| WPRIM
| ID: wpr-123227
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pneumonia
/
Streptococcus pneumoniae
/
DNA
/
RNA
/
Nasopharynx
/
Polymerase Chain Reaction
/
Reverse Transcription
/
Real-Time Polymerase Chain Reaction
/
Methods
Type of study:
Diagnostic study
/
Prognostic study
Limits:
Child
/
Humans
Language:
Korean
Journal:
Pediatric Infection & Vaccine
Year:
2016
Type:
Article
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