Evaluation of the Seeplex® Meningitis ACE Detection Kit for the Detection of 12 Common Bacterial and Viral Pathogens of Acute Meningitis
2
조회
0
다운로드
5
인용
0
참고문헌
키워드
초록
Background: Bacterial meningitis is an infectious disease with high rates of mortality and high frequency of severe sequelae. Early identification of causative bacterial and viral pathogens is important for prompt and proper treatment of meningitis and for prevention of life-threatening clinical outcomes. In the present study, we evaluated the value of the Seeplex Meningitis ACE Detection kit (Seegene Inc., Korea), a newly developed multiplex PCR kit employing dual priming oligonucleotide methods, for diagnosing acute meningitis. Methods: Analytical sensitivity of the kit was studied using reference strains for each pathogen targeted by the kit, while it’s analytical specificity was studied using the human genome DNA and 58 clinically well-identified reference strains. For clinical validation experiment, we used 27 control cerebrospinal fluid (CSF) samples and 78 clinical CSF samples collected from patients at the time of diagnosis of acute meningitis. Results: The lower detection limits ranged from 101 copies/μL to 5×101 copies/μL for the 12 viral and bacterial pathogens targeted. No cross-reaction was observed. In the validation study, high detection rate of 56.4% was obtained. None of the control samples tested positive, i.e., false-positive results were absent. Conclusions: The Seeplex Meningitis ACE Detection kit showed high sensitivity, specificity, and detection rate for the identification of pathogens in clinical CSF samples. This kit may be useful for rapid identification of important acute meningitis-causing pathogens.
Abstract
Background: Bacterial meningitis is an infectious disease with high rates of mortality and high frequency of severe sequelae. Early identification of causative bacterial and viral pathogens is important for prompt and proper treatment of meningitis and for prevention of life-threatening clinical outcomes. In the present study, we evaluated the value of the Seeplex Meningitis ACE Detection kit (Seegene Inc., Korea), a newly developed multiplex PCR kit employing dual priming oligonucleotide methods, for diagnosing acute meningitis. Methods: Analytical sensitivity of the kit was studied using reference strains for each pathogen targeted by the kit, while it’s analytical specificity was studied using the human genome DNA and 58 clinically well-identified reference strains. For clinical validation experiment, we used 27 control cerebrospinal fluid (CSF) samples and 78 clinical CSF samples collected from patients at the time of diagnosis of acute meningitis. Results: The lower detection limits ranged from 101 copies/μL to 5×101 copies/μL for the 12 viral and bacterial pathogens targeted. No cross-reaction was observed. In the validation study, high detection rate of 56.4% was obtained. None of the control samples tested positive, i.e., false-positive results were absent. Conclusions: The Seeplex Meningitis ACE Detection kit showed high sensitivity, specificity, and detection rate for the identification of pathogens in clinical CSF samples. This kit may be useful for rapid identification of important acute meningitis-causing pathogens.
참고문헌 (16)
- Grimwood K, "Twelve year outcomes following bacterial meningitis: further evidence for persisting effects", Arch Dis Child, 83, pp.111~116 (2000)
- Weisfelt M, "Clinical features, complications, and outcome in adults with pneumococcal meningitis: a prospective case series", Lancet Neurol, 5, pp.123~129 (2006)
- Bäckman A, "Evaluation of an extended diagnostic PCR assay for detection and verification of the common causes of bacterial meningitis in CSF and other biological samples", Mol Cell Probes, 13, pp.49~60 (1999)
- Schuurman T, "Prospective study of use of PCR amplification and sequencing of 16S ribosomal DNA from cerebrospinal fluid for diagnosis of bacterial meningitis in a clinical setting", J Clin Microbiol, 42, pp.734~740 (2004)
- Welinder-Olsson C, "Comparison of broad-range bacterial PCR and culture of cerebrospinal fluid for diagnosis of community-acquired bacterial meningitis", Clin Microbiol Infect, 13, pp.879~886 (2007)
- Clarke SC, "Detection of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae in blood and cerebrospinal fluid using fluorescence-based PCR", Methods Mol Biol, 345, pp.69~77 (2006)
- Smith K, "Automation of a fluorescence-based multiplex PCR for the laboratory confirmation of common bacterial pathogens", J Med Microbiol, 53, pp.115~117 (2004)
- Bøving MK, "Eight-plex PCR and liquid-array detection of bacterial and viral pathogens in cerebrospinal fluid from patients with suspected meningitis", J Clin Microbiol, 47, pp.908~913 (2009)
- Ren HY, "Detection of bacterial meningitis by multiplex polymerase chain reaction", Zhongguo Yi Miao He Mian Yi, 16, pp.47~51 (2010)
- Uzuka R, "Rapid diagnosis of bacterial meningitis by using multiplex PCR and real time PCR", Pediatr Int, 46, pp.551~554 (2004)
- Chang WN, "Changing epidemiology of adult bacterial meningitis in southern taiwan: a hospital-based study", Infection, 36, pp.15~22 (2008)
- Thwaites GE, "Diagnosis of adult tuberculous meningitis by use of clinical and laboratory features", Lancet, 360, pp.1287~1292 (2002)
- Chun JY, "Dual priming oligonucleotide system for the multiplex detection of respiratory viruses and SNP genotyping of CYP2C19 gene", Nucleic Acids Res, 35, pp.e40~ (2007)
- 성흥섭, "Rhinovirus, Human Metapneumovirus, Coronavirus 검출을 위한SeeplexTM RV Detection 키트의 평가", Annals of Laboratory Medicine, 28(2), pp.109~117 (2008), 대한진단검사의학회
- 박주원, "소아의 세균성 호흡기감염의 진단을 위한 SeeplexTM Pneumobacter Multiplex PCR 키트의 평가", Annals of Laboratory Medicine, 29(4), pp.307~313 (2009), 대한진단검사의학회
- Higgins RR, "Evaluation and verification of the Seeplex Diarrhea-V ACE assay for simultaneous detection of adenovirus, rotavirus and norovirus genogroups I and II in clinical stool specimens", J Clin Microbiol, 49, pp.3154~3162 (2011)