PMID- 30156509
OWN - NLM
STAT- MEDLINE
DCOM- 20190424
LR  - 20190424
IS  - 0374-9096 (Print)
IS  - 0374-9096 (Linking)
VI  - 52
IP  - 3
DP  - 2018 Jul
TI  - [Optimization of real-time multiplex polymerase chain reaction for the diagnosis 
      of acute bacterial meningitis and Neisseria meningitidis serogrouping].
PG  - 221-232
LID - 10.5578/mb.67019 [doi]
AB  - Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae type 
      B are responsible for about 80-85% of acute bacterial meningitis cases among 
      adults and in our country it is mandatory to report bacterial meningitis cases 
      caused by these agents. In recent years, disease control programs have focused 
      especially on these three pathogens in our country. It is very important to know 
      the causative agent of meningitis for prevention/control measures, choice of 
      treatment regimen, and prediction of the severity and the course of the disease. 
      Rapid diagnosis of meningococcal meningitis is especially vital. In addition, the 
      identification of N.meningitidis serogroups in the community is also important 
      for the decision of the type and combination of vaccine to be administered. The 
      aim of this study was to optimize the real-time multiplex polymerase chain 
      reaction (Rt-multiplex PCR) for the diagnosis of the disease and serogrouping of 
      N.meningitidis in clinical samples in a direct, quick and reliable way. In this 
      study, three Rt-multiplex PCR assays were optimized and standardized for the 
      detection of N.meningitidis, H.influenzae and S.pneumoniae (NHS mix) and for the 
      serogrouping of N.meningitidis (BCY mix and AWX mix) in our laboratory. 
      Sensitivity of the Rt-multiplex PCR method was detected by reference strains and 
      simulation studies. Forty three samples (41 cerebrospinal fluid (CSF), 1 serum 
      and 1 clinical isolate) with the suspicion of acute bacterial meningitis and six 
      nasopharyngeal isolates sent to National Reference Laboratory for Molecular 
      Microbiology between July 2012-May 2014 were included in the present study. All 
      samples were examined with the Rt-multiplex PCR methods. Clinical isolates were 
      studied by both conventional and Rt-multiplex PCR methods. Oxidase, catalase 
      test, Gram stain, API-NH and agglutination tests with specific antisera were 
      performed in the National Respiratory Pathogens Reference Laboratory. The 
      detection limit of the method, which was optimized and standardized in our 
      laboratory was determined as 102 cfu/ml. The CT (threshold cycle) value in this 
      dilution was detected approximately as 35. N.meningitidis was detected in 14 of 
      the 41 CSF samples by the NHS mix. However, only 10 of the positive samples could 
      be typed with BCY mix and AWX mix. Eight (80%) of them were serogroup B, one of 
      each was (10%) serogroup A and serogroup W135, respectively. All the isolates 
      (six nasopharyngeal and one clinical specimen) were identified as N.meningitidis 
      serogroup B by Rt-multiplex PCR and the isolates were also confirmed by 
      conventional methods. The CSF specimens with CT value > 35 could not be typed. We 
      concluded that the Rt-multiplex PCR method is a rapid and reliable test for the 
      direct diagnosis of acute bacterial meningitis due to NHS and serogrouping of 
      N.meningitidis. Rapid diagnosis plays an important role for the treatment and 
      control of the disease, and serogrouping of the agent plays an important role in 
      terms of prevention/control and vaccination policies.
FAU - Guldemir, Dilek
AU  - Guldemir D
AD  - General Directorate of Public Health Turkey, Department of Microbiology Reference 
      Laboratories, National Molecular Microbiology Reference Laboratory, Ankara, 
      Turkey.
FAU - Turan, Meral
AU  - Turan M
AD  - General Directorate of Public Health Turkey, Department of Microbiology Reference 
      Laboratories, National Respiratory Pathogens Reference Laboratory, Ankara, 
      Turkey.
FAU - Bakkaloglu, Zekiye
AU  - Bakkaloglu Z
AD  - General Directorate of Public Health Turkey, Department of Microbiology Reference 
      Laboratories, National Molecular Microbiology Reference Laboratory, Ankara, 
      Turkey.
FAU - Nar Otgun, Selin
AU  - Nar Otgun S
AD  - General Directorate of Public Health Turkey, Department of Microbiology Reference 
      Laboratories, National Respiratory Pathogens Reference Laboratory, Ankara, 
      Turkey.
FAU - Durmaz, Riza
AU  - Durmaz R
AD  - Yildirim Beyazit University Faculty of Medicine, Department of Medical 
      Microbiology, Ankara, Turkey.
LA  - tur
PT  - Journal Article
TT  - Akut bakteriyel menenjit tanisi ve Neisseria meningitidis serogruplandirmasinda 
      gercek zamanli multipleks polimeraz zincir reaksiyonunun optimizasyonu.
PL  - Turkey
TA  - Mikrobiyol Bul
JT  - Mikrobiyoloji bulteni
JID - 7503830
SB  - IM
MH  - Humans
MH  - *Meningitis, Bacterial/diagnosis
MH  - *Multiplex Polymerase Chain Reaction
MH  - *Neisseria meningitidis/classification/genetics
MH  - Real-Time Polymerase Chain Reaction
EDAT- 2018/08/30 06:00
MHDA- 2019/04/25 06:00
CRDT- 2018/08/30 06:00
PHST- 2018/08/30 06:00 [entrez]
PHST- 2018/08/30 06:00 [pubmed]
PHST- 2019/04/25 06:00 [medline]
AID - 10.5578/mb.67019 [doi]
PST - ppublish
SO  - Mikrobiyol Bul. 2018 Jul;52(3):221-232. doi: 10.5578/mb.67019.