NDLI: A fast real-time polymerase chain reaction method for sensitive and specific detection of the Neisseria gonorrhoeae porA pseudogene.

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A fast real-time polymerase chain reaction method for sensitive and specific detection of the Neisseria gonorrhoeae porA pseudogene.

Content Provider	Semantic Scholar
Author	Hjelmevoll, Stig Ove Olsen, Merethe Elise Sollid, Johanna Ulrica Ericson Haaheim, Håkon Unemo, Magnus Skogen, Vegard
Copyright Year	2006
Abstract	Ever since the advent of molecular methods, the diagnostics of Neisseria gonorrhoeae has been troubled by false negative and false positive results compared with culture. Commensal Neisseria species and Neisseria meningitidis are closely related to N. gonorrhoeae and may cross-react when using molecular tests comprising too-low specificity. We have devised a real-time polymerase chain reaction (PCR), including an internal amplification control, that targets the N. gonorrhoeae porA pseudogene. DNA was automatically isolated on a BioRobot M48. Our subsequent PCR method amplified all of the different N. gonorrhoeae international reference strains (n = 34) and N. gonorrhoeae clinical isolates (n = 176) but not isolates of the 13 different nongonococcal Neisseria species (n = 68) that we tested. Furthermore, a panel of gram-negative bacterial (n = 18), gram-positive bacterial (n = 23), fungal (n = 1), and viral (n = 4) as well as human DNA did not amplify. The limit of detection was determined to be less than 7.5 genome equivalents/PCR reaction. In conclusion, the N. gonorrhoeae porA pseudogene real-time PCR developed in the present study is highly sensitive, specific, robust, rapid and reproducible, making it suitable for diagnosis of N. gonorrhoeae infection.
Starting Page	29
Ending Page	38
Page Count	10
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://www.copanusa.com/files/8314/2712/6678/Hjelmevoll_J_Mol_Diagn_2006.pdf
PubMed reference number	17065426v1
Volume Number	8
Issue Number	5
Journal	The Journal of molecular diagnostics : JMD
Language	English
Access Restriction	Open
Subject Keyword	Equivalent Weight Molecular Genetic Technique Neisseria gonorrhoeae Neisseria meningitidis Pseudogenes Real-Time Polymerase Chain Reaction Reverse Transcriptase Polymerase Chain Reaction gram
Content Type	Text
Resource Type	Article

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