International Conference on Medical and Clinical Microbiology July 03-04, 2017 AVANI Atrium Bangkok, Bangkok, Thailand

Hasyanee Binmaeil. was born in Thailand in 1987. She is Ph.D student in Dept. of Medical Microbiology and Immunology, Faculty of Medicine at Universiti Kebangsaan Malaysia. Her research interests is on Helicobacter pylori study which focus on the development of the molecular techniques for detection of H. pylori directly in stomach of patients with dyspepsia. She have presented a poster at Postgraduate Conference and published an article in Journal of Global Veterinaria (2014).

The aim of this study was to evaluate the accuracy of this new real-time PCR (qPCR) test for detection of H. pylori infection in biopsy specimens in comparison to conventional tests: histology, RUT and culture. Patients who attending Endoscopy Unit, UKMMC with dyspepsia and undergoing oesophagogastroduodenoscopy between April 2014 to August 2015 were recruited. Stomach biopsies were collected for histology, Rapid urease test (RUT), culture and qPCR analysis. A total of 288 biopsy samples, of which 34 biopsy samples were considered positive for H. pylori infection by conventional methods (concordant positive results on 2 or more tests). The remaining 254 biopsies (88.19%) were considered negative for H. pylori. In contrast, the new real-time PCR (qPCR) test detected 64 H. pylori infection in biopsies of patients, which is significantly higher than conventional test (P<0.0001). H. pylori infection rate determined by conventional methods varied from 0.35% to 3.13% among different age groups in 288 patients with dyspepsia. H. pylori infection rate determined by qPCR method is higher and varied from 0.69% to 5.21% among different age groups. H. pylori infection rate is the highest at 51-60 and 71-80 years old as determined using qPCR and conventional methods, respectively. Results of the present study indicate that qPCR is more sensitive than conventional methods to detect H. pylori infection in patients with dyspepsia. In summary, we have developed a rapid and sensitive q-PCR method for detection of H. pylori directly from biopsy specimens. This technique offers a significant improvement over other available conventional methods for detecting H. pylori in clinical and research samples.

Michał Styczyński, student of Biotechnology in Department of Virology at University of Warsaw. He is working on human epithelium tissues such as HEC-1B and ME-180 and with bacteria from Neisseria genus. Michał is interested in LSCM and SEM microscopy, and he is already exploring structure of Neisseria gonorrhoeae biofilms. Michał is also very interested in bioremediation and application of practical biotechnologies.

INTRODUCTION:\r\nEffective pathogen invasion requires an environment which provide proper concentration of essential ions so in which bacteria population can grow up and successfully colonize.\r\nIron is a very important factor for almost all organisms around the world. It is necessary for a basic life processes such as for example DNA biosynthesis. The bioavailability of free iron ions is difficult and it is often a limiting factor for microorganism life.\r\nNeisseria gonorrhoeae is the obligatory pathogen for a human and is an etiological factor of gonorrhea disease, which is a global health problem.\r\nNeisseria sicca and Neisseria lactamica are examples of commensal bacteria, common in human upper respiratory tract. However, during immune deficiencies, they can cause diseases such as sepsis, arthritis or pneumonia.\r\nN. gonorrhoeae as well as N. sicca and N. lactamica needs iron ions for survival in the environment.\r\nMATERIALS AND METHODS:\r\nBacterial strains:\r\n Neisseria gonorrhoeae FA 1090 ATCC 700825\r\n Neisseria sicca ATCC 29256\r\n Neisseria lactamica ATCC 23970\r\nBacterial cultures were grown in liquid GC medium with different availability and amount of iron:\r\n(i) + 250 μM Fe3(NO3)3 · 9H2O\r\n(ii) + 500 μM desferal (iron ions chelator)\r\nControls were grown under standard conditions.\r\nThree times of culture incubation were tested:\r\n1h, 4h and 24h.\r\nAfter incubation, the OD600 of the culture was measured.\r\nAdditionaly, formed biofilm was stained with crystalline violet, dissolved in ethanol and OD570 was measured.\r\nRESULTS:\r\nAfter 1h and 4h, neither for iron ions and desferal there are not statistical significance in survivability and in forming biofilm. After 24h addition of desferal significantly reduced bacterial growth in all tested bacteria species.\r\nFuthermore after 24h desferal induced forming biofilm in N. gonorrhoeae ** but reduced in N. sicca and N. lactamica. Iron ions reduced forming biofilm in N. sicca** but stimulated in N. lactamica*.