【病毒外文文獻】2015 Development of a new detection tool by real time PCR for the detection of Middle East Respiratory Syndrome human Co

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Abstracts Journal of Clinical Virology 70 2015 S1 S126 S59 Abstract No 1683 Presentation at ESCV 2015 Poster 1 Frequent detection of human papillomavirus DNA in oral lesions A Pierangeli F Cannella U Romeo G Palaia A Polimeni G Antonelli Sapienza University Rome Italy Background Human papillomavirus HPV is estimated to be the cause of 40 80 of oropharyngeal squamous cell carcinoma OSCC The incidence of OSCC has significantly increased in the last decade and the prevalence of oral HPV infection in the general population augmented as well Therefore there is a need to evalu ate the effectiveness of HPV detection for OSCC prevention using methods less invasive than surgical biopsy We sought to detect HPV DNA in oral brushings from oral potentially malignant disor ders OPMD and oral carcinomas HPV DNA positivity viral loads and the HPV16 integrational status were evaluated in relation to clinical data Methods A total of 134 individuals attending Odontostoma tologic Clinics were enrolled 77 patients affected by benign oral lesions OPMD or oral carcinoma and 57 controls with no lesion in the oral mucosa Oral cells were collected using Cytobrush a not invasive but site specific method HPV DNA was detected with quantitative real time PCR qPCR for the more common low risk HPV 6 11 and high risk 16 18 31 33 53 58 genotypes Results In patients group 40 77 51 9 samples were HPV positive a rate significantly higher than that found in the con trols 33 3 The percentage of high risk HPVs and HPV DNA loads in samples from cases were significantly higher than in controls among lesions lichen planus had the highest HPV positivity rate 71 4 hairy leukoplakia the lowest 33 3 Interestingly HPV16 resulted the most frequent genotype in all types of lesions but was integrated only in 3 out of 4 HPV positive carcinomas and in 3 14 HPV positive lichen planus Conclusion This study implemented a non invasive sample collection method with a sensitive and quantitative molecular method detecting high rates of oral HPV DNA particularly in lichen planus and high integration rates in HPV positive carcinomas The fact that HPV oral infection may be common suggest the utility of viral load and integrational status determination to discriminate those lesions at higher risk of malignant progression http dx doi org 10 1016 j jcv 2015 07 139 Abstract No 1687 Presentation at ESCV 2015 Poster 1 Development of a new detection tool by real time PCR for the detection of Middle East Respiratory Syndrome human Coronavirus MERS HCoV combining specific primers probe and a RNA internal control ready to use premix M Vignoles 1 P Marechal 1 M Dube 1 C Barranger 1 D Heckel 2 M Joannes 1 1 bioM rieux Verniolle France 2 bioM rieux Grenoble France Background Since the emergence of Middle East Respira tory Syndrome Coronavirus MERS CoV in 2012 in the Arabian Peninsula many questions remain on modes of transmission and sources of virus The MERS HCoV causes severe respiratory illness The epidemic origins are uncertain but it is probably a zoonosis The reservoirs could be the bat or the camel In 2015 04 29 the MERS HCoV is responsible of 1110 laboratory confirmed cases and 442 deaths WHO In outbreak situations especially with emerg ing organisms causing severe human diseases it is important to develop quickly a test to detect the virus involved bioM rieux developed a real time PCR assay for the rapid detection of MERS HCoV combining specific primers probe and a RNA internal control ready to use premix The results of the analytical sensitivity the precision determination and the exclusivity study are presented Methods A set of primers and probe Mers hCoV primers r gene 20 010 and Mers hCoV probe r gene 20 011 bioM rieux were designed on the S gene coding for the spike structural protein This set was evaluated in combination with a ready to use premix for RNA internal control to constitute a duplex RT PCR assay The internal control added before the extrac tion step allows to check simultaneously extraction efficiency and presence of inhibitors Extractions were performed on NucliSENS easyMAG bioM rieux followed by amplifications on 7500 Fast Real Time PCR System Applied Biosystems The analytical sen sitivity was performed on 20 replicates of a wide dilution range of In Vitro transcripts The precision determination was determined on 18 replicates of 3 points corresponding at 40 20 and 8 times LoD 95 The exclusivity study was carried out on major human respiratory viruses Results The analytical sensitivity was determined by Probit analysis MiniTab The LoD 95 hit rate detection was 2 89 log copies mL of In Vitro transcripts The coefficients of variation obtained in the precision test were ranged between 1 28 and 2 26 for the MERS HCoV and between 1 95 and 2 39 for the inter nal control The exclusivity study was performed with the major human respiratory viruses including other human coronaviruses and no cross reaction was observed Conclusion The high quality in analytical sensitivity 2 89 and robustness 1 28 2 39 were demonstrated The combination of the MersCoV primers and probe in association with the RNA inter nal control ready to use premix provides a useful tool for the detection of MERS HCoV virus http dx doi org 10 1016 j jcv 2015 07 140 Abstract No 1690 Presentation at ESCV 2015 Poster 1 Acute flaccid paralysis surveillance system in Norway detected two cases of enterovirus D68 infection N Milhano 1 K Bragstad 2 H C Pfeiffer 3 K Vainio 2 J Bj rnholt 2 A M B Kran 4 S Dudman 5 1 Department of Virology Norwegian Institute of Public Health European Public Health Microbiology Training Programme Norway 2 Norwegian Institute of Public Health Norway 3 Department of Pediatrics Oslo University Hospital Norway 4 Department of Microbiology Oslo University Hospital University of Oslo Institute of Clinical Medi Norway 5 National Institute of Health Oslo Norway Background WHO recommends surveillance of patients with acute flaccid paralysis AFP to ensure the eradication of wild poliovirus In Norway the AFP surveillance is used as the gold standard for polio surveillance with supplementary enterovirus surveillance Here we describe the Norwegian AFP surveillance sys tem as well as its performance by assessing the frequency of AFP