Detection and Quantification of Human Adenoviruses in Surface Waters by Nested PCR, TaqMan Real-Time PCR and Cell Culture Assays

2007 ◽  
Vol 191 (1-4) ◽  
pp. 83-93 ◽  
Author(s):  
M. Muscillo ◽  
M. Pourshaban ◽  
M. Iaconelli ◽  
S. Fontana ◽  
A. Di Grazia ◽  
...  
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
Surface Waters ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Human Adenoviruses ◽  
Detection And Quantification

scholarly journals Comparison of real-time PCR protocols in detection and quantification of fruit tree 16SrX group phytoplasmas

Genetika ◽  
2016 ◽  
Vol 48 (2) ◽  
pp. 629-642 ◽  
Author(s):  
Tomas Kiss ◽  
Tomas Necas ◽  
Jana Necasova
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Fruit Tree ◽  
Amplification Efficiency ◽  
Specificity And Sensitivity ◽  
Intercalating Dye ◽  
Detection Characteristics ◽  
Detection And Quantification

In this work, two real-time PCR protocols based on intercalating dye and two on hydrolysis probes were tested using field collected fruit tree samples infected by 16SrX group (AP, PD and ESFY) phytoplasmas. Specificity and sensitivity of protocols and amplification efficiency were the main testing parameters. Results of real-time PCR protocols were compared to nested PCR. All real-time PCR protocols confirmed their specificity of detection. All real-time PCR protocols were 10-100 times more sensitive than nested PCR. Afterall real-time PCR protocols based on hydrolysis probes were 10 times more sensitive than protocols based on intercalating dyes. Among protocols based on hydrolysis probes, slightly better detection characteristics were shown by protocol by CHRISTENSEN et al. (2004).

Identifying the etiologic role of Parvovirus B19 in non-immune hydrops fetalis by histopathology, immunohistochemistry and nucleic acid testing: a retrospective study

Open Medicine ◽  
2007 ◽  
Vol 2 (3) ◽  
pp. 271-279 ◽  
Author(s):  
Koray Ergunay ◽  
Gulcin Altinok ◽  
Bora Gurel ◽  
Ahmet Pinar ◽  
Arzu Sungur ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
San Francisco ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Parvovirus B19 ◽  
Etiologic Agent ◽  
Hydrops Fetalis ◽  
Nucleic Acid Detection

AbstractIntrauterine Parvovirus B19 infections may cause fetal anemia, non-immune hydrops fetalis or abortion. This study focuses on the pathogenic role of Parvovirus B19 in non-immune hydrops fetalis at Hacettepe University, a major reference hospital in Turkey. Twenty-two cases of non-immune hydrops fetalis were retrospectively selected out of a total of 431 hydrops fetalis specimens from the Department of Pathology archieves. Paraffine embedded tissue sections from placental and liver tissues from each case were evaluated by histopathology, immunohistochemistry, nested PCR and commercial quantitative Real-time PCR. Viral DNA was detected in placental tissues by Real-time PCR in 2 cases (2/22, 9.1%) where histopathology also revealed changes suggestive of Parvovirus B19 infection. No significant histopathologic changes were observed for the remaining sections. Nested PCR that targets the VP1 region of the viral genome and immunohistochemistry for viral capsid antigens were negative for all cases. As a result, Parvovirus B19 is identified as the etiologic agent for the development of non-immune hydrops fetalis for 9.1% of the cases in Hacettepe University, Turkey. Real-time PCR is observed to be an effective diagnostic tool for nucleic acid detection from paraffine embedded tissues. Part of this study was presented as a poster at XIIIth International Congress of Virology, San Francisco, USA (Abstract V-572).

scholarly journals Application of quantitative real-time PCR compared to filtration methods for the enumeration of Escherichia coli in surface waters within Vietnam

2016 ◽  
Vol 15 (1) ◽  
pp. 155-162 ◽  
Author(s):  
Pierangeli G. Vital ◽  
Nguyen Thi Van Ha ◽  
Le Thi Hong Tuyet ◽  
Kenneth W. Widmer
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Surface Waters ◽  
Real Time Pcr ◽  
Membrane Filtration ◽  
Quantitative Real Time Pcr ◽  
Wet Season ◽  
Culture Methods ◽  
Fecal Indicator ◽  
Filtration Method

Surface water samples in Vietnam were collected from the Saigon River, rural and suburban canals, and urban runoff canals in Ho Chi Minh City, Vietnam, and were processed to enumerate Escherichia coli. Quantification was done through membrane filtration and quantitative real-time polymerase chain reaction (PCR). Mean log colony-forming unit (CFU)/100 ml E. coli counts in the dry season for river/suburban canals and urban canals were log 2.8 and 3.7, respectively, using a membrane filtration method, while using Taqman quantitative real-time PCR they were log 2.4 and 2.8 for river/suburban canals and urban canals, respectively. For the wet season, data determined by the membrane filtration method in river/suburban canals and urban canals samples had mean counts of log 3.7 and 4.1, respectively. While mean log CFU/100 ml counts in the wet season using quantitative PCR were log 3 and 2, respectively. Additionally, the urban canal samples were significantly lower than those determined by conventional culture methods for the wet season. These results show that while quantitative real-time PCR can be used to determine levels of fecal indicator bacteria in surface waters, there are some limitations to its application and it may be impacted by sources of runoff based on surveyed samples.

scholarly journals Detection of enterovirus in mussels from Morocco by cell culture and real-time PCR

2017 ◽  
Vol 16 (34) ◽  
pp. 1791-1799 ◽  
Author(s):  
Meryem Idrissi Azzouzi Lalla ◽  
Senouci Samira ◽  
El Qazoui Maria ◽  
Oumzil Hicham ◽  
Naciri Mariam
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
Real Time Pcr

scholarly journals Detection and quantification of Streptococcus pneumoniae from Iranian patients with pneumonia and individual carriers by real time PCR

2011 ◽  
Vol 10 (60) ◽  
pp. 12826-12832 ◽  
Author(s):  
Nomanpour Bizhan ◽  
Ghodousi Arash ◽  
Babaei Toraj ◽  
AliJavad Mousavi Seyd ◽  
Asadi Soroor ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
Iranian Patients ◽  
Detection And Quantification

scholarly journals Detection of Peanut Allergen Traces with a Real Time PCR Assay - The Challenge to Protect Food-Allergic Consumers

2017 ◽  
Vol 7 (1) ◽  
pp. 32 ◽  
Author(s):  
Dimitra Houhoula ◽  
Stamatios Koussissis ◽  
Vladimiros Lougovois ◽  
John Tsaknis ◽  
Dimitra Kassavita ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Food Products ◽  
Molecular Techniques ◽  
Food Allergens ◽  
Pcr Assay ◽  
Peanut Allergen ◽  
Pcr Method ◽  
Detection And Quantification

The aim of the present study was the implementation of molecular techniques in the detection and quantification of allergic substances of peanut in various kinds of food products, e.g., breakfast cereals, chocolates and biscuits that are frequently related to allergies. In some cases, the presence of peanuts can be due to contamination during production and are not declared on the label. A total of 152 samples were collected from supermarkets and were analysed by a Real Time PCR method. The results indicated that 125 samples (83,3%) were found positive in peanut traces but the most important finding is that from the 84 samples that had no allergen declaration for peanuts, 48 (57,1%) of them were found positive. In conclusion, Real Time PCR can be a very important tool for the rapid detection and quantification of food allergens.

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