Hettich EBA 12 Centrifuge Used in Neisseria Meningitidis Detection Study
A study entitled “Evaluation of touch-down real-time PCR based on SYBR Green I ﬂuorescent dye for the detection of Neisseria meningitidis in clinical samples” was published in the FEMS Immunology and Medical Microbiology journal in December 2004. The study used the Hettich EBA 12 centrifuge to compare the effectiveness of conventional polymerase chain reaction (PCR) against real-time polymerase chain reaction (RTPCR) in the detection of meningococcal DNA in lab samples.
According to the study, Neisseria meningitidis is a type of bacteria that causes childhood septicaemia and meningitis. It is also among the leading infectious causes of death in healthy children and young adults in developed countries. In the past decade, detection methods based on PCR for meningococcal DNA have proven to be valuable for the diagnosis of invasive meningococcal disease (IMD).
For the experiment, samples from 245 individuals in different areas of Greece were collected between 2002 and 2003. 273 samples taken from CSF, blood, and bacterial isolates were then tested with both conventional and real-time PCR assays. The samples were first centrifuged at 1700g with the Hettich EBA 12 centrifuge for 10 minutes to isolate target DNA from CSF. They were later heated at 100 °C for 30 minutes and centrifuged for a second time at 10,000g for 8 minutes.
Results of the study showed that the real-time PCR assay was effective in detecting meningococcal DNA rapidly and accurately. The RTPCR was able to distinguish N. meningitidis from N. lactamica DNA in two samples, of which the conventional PCR gave a false positive result. It was concluded that the RTPCR would be able to provide a convenient and reproducible system with minimal contamination risks. Additionally, the RTPCR could also serve as a replacement for conventional assays in laboratories using PCR for non-culture diagnosis of IMD.
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