Extraction of DNA from for the Real-Time PCR Detection of Mycobacterium ulcerans

2021 ◽  
pp. 63-69
Author(s):  
Janet A. M. Fyfe ◽  
Caroline J. Lavender
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Detection ◽  
Mycobacterium Ulcerans ◽  
The Real

Combined effects of matrix and gene marker on the real-time PCR detection of wheat

2016 ◽  
Vol 51 (7) ◽  
pp. 1680-1688 ◽  
Author(s):  
Begoña Martín-Fernández ◽  
Joana Costa ◽  
Maria Beatriz Prior Pinto Oliveira ◽  
Beatriz López-Ruiz ◽  
Isabel Mafra
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Detection ◽  
Gene Marker ◽  
Combined Effects ◽  
The Real

Enhanced enrichment and detection of thermotolerant Campylobacter species from water using the Portable Microbe Enrichment Unit and real-time PCR

2009 ◽  
Vol 55 (7) ◽  
pp. 849-858 ◽  
Author(s):  
Tarja Pitkänen ◽  
Juliane Bräcker ◽  
Ilkka T. Miettinen ◽  
Anneli Heitto ◽  
Jouni Pesola ◽  
...  
Keyword(s):  
Drinking Water ◽  
Real Time ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Pcr Detection ◽  
Detection Time ◽  
The Real ◽  
International Standard ◽  
Campylobacter Species

An enhanced enrichment using the Portable Microbe Enrichment Unit (PMEU) with the microaerobic bubbling of broths was applied for the detection of thermotolerant Campylobacter species from water. This PMEU enrichment was compared with the conventional static enrichment of the international standard ISO 17995:2005. In addition, Campylobacter detection after enrichment using a real-time PCR detection was compared with colony counts. The tests with stressed Campylobacter jejuni cells in drinking water indicated that the PMEU enrichment yielded a significantly higher number of Campylobacter cells in the Bolton broth compared with the conventional static incubation. Application of the real-time PCR technique shortened the Campylobacter detection time. This combination of method modifications can be used for Campylobacter detection from water and adds methodological repertoire for the rapid survey and management of waterborne outbreaks.

scholarly journals Rectal Swabs as an Alternative Sample Collection Method to Bulk Stool for the Real-Time PCR Detection of Giardia duodenalis

2020 ◽  
Vol 103 (3) ◽  
pp. 1276-1282 ◽  
Author(s):  
Jacqueline R. M. A. Maasch ◽  
Ahmed M. Arzika ◽  
Catherine Cook ◽  
Elodie Lebas ◽  
Nils Pilotte ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Giardia Duodenalis ◽  
Pcr Detection ◽  
Sample Collection ◽  
Collection Method ◽  
The Real ◽  
Rectal Swabs

The Establishment of Real-Time PCR Detection System of Aspergillus Niger in Peanut

2016 ◽  
Vol 835 ◽  
pp. 333-337
Author(s):  
Chu Shu Zhang ◽  
Jie Sun ◽  
Li Na Yu ◽  
Jie Bi ◽  
Wei Wei Zhao
Keyword(s):  
Aspergillus Niger ◽  
Real Time ◽  
Correlation Coefficient ◽  
Real Time Pcr ◽  
Detection System ◽  
Dna Detection ◽  
Pcr Primers ◽  
Pcr Detection ◽  
The Real

In this study, a set of real-time PCR primers for the detection of aflatoxin in peanut was successfully designed according to the AFLR gene of Aspergillus niger. Upstream primer was 5'-AACCTGATGACGACTGATAT-3'; Downstream primer was 5'-AGCACCTTGAGAACGATAA-3'; The real-time PCR detection system established in this study had good specificity, and the sensitivity of DNA detection can reach 3×10-13 ng/ul. The correlation coefficient between the contents of aflatoxin in peanut samples and PCR Ct of Real-time was-0.718, and the correlation coefficient was significantly negative.

APPLICATION OF REAL-TIME PRC TECHNIQUE AND REVERSE DOT-BLOT FOR DETECTION THE HIGH RISK TYPES OF HUMAN PAPILLOMAVIRUS CAUSING CERVICAL CANCER

2017 ◽  
pp. 99-103
Author(s):  
Van Bao Thang Phan ◽  
Hoang Bach Nguyen ◽  
Van Thanh Nguyen ◽  
Thi Nhu Hoa Tran ◽  
Viet Quynh Tram Ngo
Keyword(s):  
Cervical Cancer ◽  
High Risk ◽  
Real Time ◽  
Real Time Pcr ◽  
Dot Blot ◽  
The Real ◽  
Dot Blot Assay ◽  
Hpv Types ◽  
High Risk Hpv ◽  
Reverse Dot Blot

Introduction: Infection with HPV is the main cause of cervical cancer. Determining HPV infection and the types of HPV plays an important role in diagnosis, treatment and prognosis of cervicitis/cervical cancer. Aims: Determining proportion of high-risk HPV types and the occurrence of coinfection with multiple HPV types. Methods: 177 women with cervicitis or abnormal Pap smear result were enrolled in the study. Performing the real-time PCR for detecting HPV and the reverse DOT-BLOT assay for determining type of HPV in cases of positive PCR. Results: 7 types of high-risk HPV was dectected, the majority of these types were HPV type 18 (74.6%) and HPV type 16 (37.6%); the proportion of infection with only one type of HPV was 30.4% and coinfection with multiple HPV types was higher (69.6%), the coinfected cases with 2 and 3 types were dominated (32.2% and 20.3%, respectively) and the coinfected cases with 4 and 5 types were rare. Conclusion: Use of the real-time PCR and reverse DOT-BLOT assay can determine the high-risk HPV types and the occurrence of coinfection with multiple HPV types. Key words: HPV type, Reverse DOT-BLOT, real-time PCR,PCR, cervical cancer

scholarly journals Real-Time PCR as an Alternative Technique for Detection of Dermatophytes in Cattle Herds

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1662
Author(s):  
Dominik Łagowski ◽  
Sebastian Gnat ◽  
Aneta Nowakiewicz ◽  
Aleksandra Trościańczyk
Keyword(s):  
Light Microscopy ◽  
Real Time ◽  
Real Time Pcr ◽  
Direct Analysis ◽  
Carrier Status ◽  
The Real ◽  
Detection Techniques ◽  
Direct Light ◽  
Microscopy Analysis ◽  
Cattle Herds

Dermatophytes are filamentous fungi with the ability to digest and grow on keratinized substrates. The ongoing improvements in fungal detection techniques give new scope for clinical implementations in laboratories and veterinary clinics, including the monitoring of the disease and carrier status. The technologically advanced methods for dermatophyte detection include molecular methods based on PCR. In this context, the aim of this study was to carry out tests on the occurrence of dermatophytes in cattle herds using qPCR methods and a comparative analysis with conventional methods. Each sample collected from ringworm cases and from asymptomatic cattle was divided into three parts and subjected to the real-time PCR technique, direct light microscopy analysis, and culture-based methods. The use of the real-time PCR technique with pan-dermatophyte primers detected the presence of dermatophytes in the sample with a 10.84% (45% vs. 34.17%) higher efficiency than direct analysis with light microscopy. Moreover, a dermatophyte culture was obtained from all samples with a positive qPCR result. In conclusion, it seems that this method can be used with success to detect dermatophytes and monitor cowsheds in ringworm cases and carriers in cattle.

scholarly journals Persistence of SARS-CoV-2 Viral RNA in Nasopharyngeal Swabs after Death: An Observational Study

2021 ◽  
Vol 9 (4) ◽  
pp. 800
Author(s):  
Francesca Servadei ◽  
Silvestro Mauriello ◽  
Manuel Scimeca ◽  
Bartolo Caggiano ◽  
Marco Ciotti ◽  
...  
Keyword(s):  
Viral Load ◽  
Observational Study ◽  
Real Time ◽  
Real Time Pcr ◽  
Pcr Detection ◽  
Viral Rna ◽  
Clinical Documentation ◽  
Post Mortem ◽  
Medical Assistance ◽  
Time Points

The aim of this study was to investigate the persistence of SARS-CoV-2 in post-mortem swabs of subjects who died from SARS-CoV-2 infection. The presence of the virus was evaluated post-mortem from airways of 27 SARS-CoV-2 positive patients at three different time points (T1 2 h; T2 12 h; T3 24 h) by real-time PCR. Detection of antibodies to SARS-CoV-2 was performed by Maglumi 2019-nCoV IgM/IgG chemiluminescence assay. SARS-CoV-2 viral RNA was still detectable in 70.3% of cases within 2 h after death and in 66,6% of cases up to 24 h after death. Our data showed an increase of the viral load in 78,6% of positive individuals 24 h post-mortem (T3) in comparison to that evaluated 2 h after death (T1). Noteworthy, we detected a positive T3 post-mortem swab (24 h after death) from 4 subjects who were negative at T1 (2 h after death). The results of our study may have an important value in the management of deceased subjects not only with a suspected or confirmed diagnosis of SARS-CoV-2, but also for unspecified causes and in the absence of clinical documentation or medical assistance.

scholarly journals Satellite DNA as a target for TaqMan real-time PCR detection of the pinewood nematode, Bursaphelenchus xylophilus

2007 ◽  
Vol 8 (6) ◽  
pp. 803-809 ◽  
Author(s):  
CECILE FRANÇOIS ◽  
CHANTAL CASTAGNONE ◽  
NEIL BOONHAM ◽  
JENNY TOMLINSON ◽  
REBECCA LAWSON ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Satellite Dna ◽  
Bursaphelenchus Xylophilus ◽  
Pcr Detection ◽  
Pinewood Nematode

Comparison of a Real-Time PCR Method with a Culture Method for the Detection of Salmonella enterica Serotype Enteritidis in Naturally Contaminated Environmental Samples from Integrated Poultry Houses

2012 ◽  
Vol 75 (4) ◽  
pp. 743-747 ◽  
Author(s):  
BWALYA LUNGU ◽  
W. DOUGLAS WALTMAN ◽  
ROY D. BERGHAUS ◽  
CHARLES L. HOFACRE
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Salmonella Enteritidis ◽  
Culture Method ◽  
Pcr Assay ◽  
Culture Methods ◽  
The Real ◽  
Selective Enrichment ◽  
Conventional Culture ◽  
Field Samples

Conventional culture methods have traditionally been considered the “gold standard” for the isolation and identification of foodborne bacterial pathogens. However, culture methods are labor-intensive and time-consuming. A Salmonella enterica serotype Enteritidis–specific real-time PCR assay that recently received interim approval by the National Poultry Improvement Plan for the detection of Salmonella Enteritidis was evaluated against a culture method that had also received interim National Poultry Improvement Plan approval for the analysis of environmental samples from integrated poultry houses. The method was validated with 422 field samples collected by either the boot sock or drag swab method. The samples were cultured by selective enrichment in tetrathionate broth followed by transfer onto a modified semisolid Rappaport-Vassiliadis medium and then plating onto brilliant green with novobiocin and xylose lysine brilliant Tergitol 4 plates. One-milliliter aliquots of the selective enrichment broths from each sample were collected for DNA extraction by the commercial PrepSEQ nucleic acid extraction assay and analysis by the Salmonella Enteritidis–specific real-time PCR assay. The real-time PCR assay detected no significant differences between the boot sock and drag swab samples. In contrast, the culture method detected a significantly higher number of positive samples from boot socks. The diagnostic sensitivity of the real-time PCR assay for the field samples was significantly higher than that of the culture method. The kappa value obtained was 0.46, indicating moderate agreement between the real-time PCR assay and the culture method. In addition, the real-time PCR method had a turnaround time of 2 days compared with 4 to 8 days for the culture method. The higher sensitivity as well as the reduction in time and labor makes this real-time PCR assay an excellent alternative to conventional culture methods for diagnostic purposes, surveillance, and research studies to improve food safety.

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