Diagnosis of Meningococcal Infection Using Internally Controlled Multiplex Real-Time PCR

AbstractMeningococcal infection requires a fast and accurate diagnostic method in order to correctly initiate the antibiotic therapy. The aim of our study was to assess the efficiency of Real Time PCR -Taq Man using sod C gene / N. meningitidis in comparison with the classical methods for the diagnosis of meningococcal infection - direct microscopy, cultivation, latex agglutination and blood culture. We have detected 24/44 (54.54%) patients with meningococcal infection. In both cases of patients with / without previous antibiotic therapy before admission, the AUC (area under curve) had the highest values for RT PCR in CSF and blood analysis. This sod C RT-PCR assay is a highly sensitive and specific method for detection of Neisseria meningitis and it would be useful to include this method like a multiplex in routine testing of patients with clinical meningococcal infection for other etiological agents also.

Download Full-text

A novel porA-based real-time PCR for detection of meningococcal carriage

Journal of Medical Microbiology ◽

10.1099/jmm.0.45847-0 ◽

2005 ◽

Vol 54 (5) ◽

pp. 463-466 ◽

Cited By ~ 24

Author(s):

J Zoe Jordens ◽

John E Heckels

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Large Scale ◽

Taqman Assay ◽

Meningococcal Infection ◽

Specific Method ◽

The Novel ◽

Wide Range ◽

Taqman Pcr ◽

Neisseria Sicca

Real-time PCR based on the capsule transfer gene (ctrA) is a significant aid in the diagnosis of meningococcal infection but fails to detect a high proportion (60 %) of non-groupable strains associated with nasopharyngeal carriage. This study aimed to design a novel real-time (TaqMan) PCR that would detect more strains of meningococci and be suitable for large-scale carriage studies. Primer and probe sequences were based on the meningococcal porA gene and designed specifically to exclude the highly related porA pseudogene in Neisseria gonorrhoeae. The specificity of the assay was confirmed by testing strains of N. gonorrhoeae known to contain the porA pseudogene together with commensal strains of Neisseria lactamica and Neisseria sicca. None of these was detected in the assay. Neisseria meningitidis strains representing a wide range of serogroups together with non-groupable strains isolated from the nasopharynx were tested by ctrA assay and the novel porA-based TaqMan PCR. All carriage strains were detected by the porA-based assay including four that gave weak or no reaction with the ctrA assay. Comparison of ctrA and porA assays on 71 throat swabs obtained from university students showed that the porA assay detected meningococcal DNA in all samples that were ctrA positive plus three that were ctrA negative but culture positive. This novel porA-based TaqMan assay provides a highly specific method for detecting meningococcal DNA that is more sensitive than the ctrA assay for detecting meningococcal carriage and is particularly suitable for carriage studies where non-groupable strains and other Neisseria are present.

Download Full-text

Feldvalidierung einer real-time PCR zum Nachweis von Mycoplasma hyopneumoniae in Nasentupfermaterial von lebenden Schweinen

Schweizer Archiv für Tierheilkunde ◽

10.1024/0036-7281.147.9.373 ◽

2005 ◽

Vol 147 (9) ◽

pp. 373-379 ◽

Cited By ~ 9

Author(s):

F. Zeeh ◽

P. Kuhnert ◽

R. Miserez ◽

M. G. Doherr ◽

W. Zimmermann

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Mycoplasma Hyopneumoniae

Download Full-text

Development of a novel real-time PCR assay for the quantitation of HBV DNA

Journal of Hepatology ◽

10.1016/s0168-8278(01)80467-0 ◽

2001 ◽

Vol 34 (0) ◽

pp. 129

Author(s):

D Paraskevis

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Hbv Dna ◽

Pcr Assay

Download Full-text

Nutzen des hoch-sensitiven real-time-PCR HBV Tests (TaqMan) zur Prädiktion der Resistenzentwicklung bei Lamivudin-Langzeittherapie

Zeitschrift für Gastroenterologie ◽

10.1055/s-0030-1264031 ◽

2010 ◽

Vol 48 (08) ◽

Author(s):

A Brodzinski ◽

F van Bömmel ◽

B Fülöp ◽

B Schlosser ◽

M Biermer ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr

Download Full-text

Nachweis des porzinen Circovirus Typ 2 und des Torque-Teno-Sus-Virus 1 und 2 in Samenproben von Ebern einer österreichischen Besamungsstation

Tierärztliche Praxis Ausgabe G Großtiere / Nutztiere ◽

10.1055/s-0038-1623060 ◽

2011 ◽

Vol 39 (04) ◽

pp. 201-204

Author(s):

A. Griessler ◽

E. Pirker ◽

H. Söllner ◽

J. Segalés ◽

T. Kekarainen ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Klinische Relevanz ◽

Torque Teno Sus Virus

Zusammenfassung Gegenstand und Ziel: Das porzine Circovirus Typ 2 (PCV-2) und das Torque-teno-Sus-Virus (TTSuV) sind in schweineproduzierenden Ländern häufig nachzuweisen. Beide Erreger können sowohl horizontal als auch vertikal übertragen werden und Ebersamen könnte ein wichtiges Übertragungsmedium darstellen. Ziel der Studie war die Abklärung der Prävalenz dieser beiden Viren in Samenproben von Ebern. Material und Methoden: Von 100 Ebern einer Besamungsstation wurde jeweils eine Samenprobe mittels quantitativer Real-Time-PCR auf PCV-2 und mittels konventioneller PCR auf TTSuV-1 und TTSuV-2 untersucht. Ergebnisse: Nur bei einem Eber der Rasse Piétrain war ein positives PCV-2-Resultat festzustellen. TTSuV-1 ließ sich in vier Samenproben, TTSuV-2 in fünf Proben nachweisen. Ein Eber wies eine Koinfektion mit beiden TTSuV-Genotypen auf. Alle TTSuV-positiven Proben stammten von Piétrain-Ebern. Schlussfolgerung und klinische Relevanz: In der vorliegenden Studie wurde erstmals in Österreich TTSuV im Samen nachgewiesen. Die Prävalenz sowohl von TTSuV als auch von PCV-2 war gering. Die klinische Relevanz einer gleichzeitigen Kontamination des Samens mit beiden Viren ist nicht klar.

Download Full-text