scholarly journals Comparison of a New Quantitative ompA-Based Real-Time PCR TaqMan Assay for Detection of Chlamydia pneumoniae DNA in Respiratory Specimens with Four Conventional PCR Assays

2003 ◽  
Vol 41 (2) ◽  
pp. 592-600 ◽  
Author(s):  
P. Apfalter ◽  
W. Barousch ◽  
M. Nehr ◽  
A. Makristathis ◽  
B. Willinger ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Chlamydia Pneumoniae ◽  
Taqman Assay ◽  
Conventional Pcr ◽  
Pcr Assays ◽  
Respiratory Specimens

Establishment of conventional PCR and real-time PCR assays for accurate, rapid and quantitative authentication of four mistletoe species

2020 ◽  
Vol 176 ◽  
pp. 112400
Author(s):  
Wook Jin Kim ◽  
Sungyu Yang ◽  
Goya Choi ◽  
Inkyu Park ◽  
Pureum Noh ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Conventional Pcr ◽  
Pcr Assays

scholarly journals Digital PCR Technology for Detection of Palm-Infecting Phytoplasmas Belonging to Group 16SrIV that Occur in Florida

Plant Disease ◽  
2018 ◽  
Vol 102 (5) ◽  
pp. 1008-1014 ◽  
Author(s):  
Brian W. Bahder ◽  
Ericka E. Helmick ◽  
De-Fen Mou ◽  
Nigel A. Harrison ◽  
Robert Davis
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Plant Pathogens ◽  
Rapid Determination ◽  
Disease Spread ◽  
Taqman Assay ◽  
Insect Vector ◽  
Palm Trees ◽  
Pcr Assays

Phytoplasmas are an economically important group of plant pathogens that negatively impact a wide variety of plants in agricultural and natural ecosystems. In Florida, palm trees are essential elements in the nursery and landscaping industries that suffer from diseases caused by phytoplasmas that are related to each other but are classified in two different subgroups, 16SrIV-A and 16SrIV-D. In this study, a TaqMan assay was developed for digital polymerase chain reaction (dPCR) to detect both palm-infecting phytoplasmas found in Florida. When compared with real-time PCR assays and nested PCR assays, dPCR was capable of detecting the phytoplasmas at much lower concentrations than was possible by using real-time PCR and nested PCR. Additionally, the assay was capable of detecting 16SrIV-B phytoplasma as well as isolates representing the 16SrI and 16SrIII phytoplasma groups. Due to sequence identity of primer annealing regions across diverse phytoplasmas, the assay is likely to be successful for detection of a wide variety of phytoplasmas. The increased sensitivity of this dPCR assay over real-time PCR will allow for earlier detection of phytoplasma infection in palm trees, as well as for screening of salivary glands of candidate insect vector species. These advantages should aid timely management decisions to reduce disease spread and rapid determination of phytoplasma transmission by vectors.

scholarly journals Development of conventional PCR and real‐time PCR assays to discriminate the origins of Chinese pepper oil and herbal materials from Zanthoxylum

2018 ◽  
Vol 99 (4) ◽  
pp. 2021-2029 ◽  
Author(s):  
Wook Jin Kim ◽  
Sungyu Yang ◽  
Goya Choi ◽  
Inkyu Park ◽  
Pureum Noh ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Conventional Pcr ◽  
Pcr Assays

scholarly journals Development of Conventional and Real-Time PCR Assays for the Rapid Detection of Group B Streptococci

2000 ◽  
Vol 46 (3) ◽  
pp. 324-331 ◽  
Author(s):  
Danbing Ke ◽  
Christian Ménard ◽  
François J Picard ◽  
Maurice Boissinot ◽  
Marc Ouellette ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Culture Method ◽  
Conventional Pcr ◽  
Pcr Assay ◽  
Group B Streptococci ◽  
Standard Culture ◽  
Pcr Assays ◽  
Group B

Abstract Background: Group B streptococci (GBS), or Streptococcus agalactiae, are the leading bacterial cause of meningitis and bacterial sepsis in newborns. Currently available rapid methods to detect GBS from clinical specimens are unsuitable for replacement of culture methods, mainly because of their lack of sensitivity. Methods: We have developed a PCR-based assay for the rapid detection of GBS. The cfb gene encoding the Christie-Atkins-Munch-Petersen (CAMP) factor was selected as the genetic target for the assay. The PCR primers were initially tested by a conventional PCR method followed by gel electrophoresis. The assay was then adapted for use with the LightCyclerTM. For this purpose, two fluorogenic adjacent hybridization probes complementary to the GBS-specific amplicon were designed and tested. In addition, a rapid sample-processing protocol was evaluated by colony-forming unit counting and PCR. A total of 15 vaginal samples were tested by both standard culture method and the two PCR assays. Results: The conventional PCR assay was specific because it amplified only GBS DNA among 125 bacterial and fungal species tested, and was able to detect all 162 GBS isolates from various geographical areas. This PCR assay allowed detection of as few as one genome copy of GBS. The real-time PCR assay was comparable to conventional PCR assay in terms of sensitivity and specificity, but it was more rapid, requiring only ∼30 min for amplification and computer-based data analysis. The presence of vaginal specimens had no detrimental effect on the sensitivity of the PCR with the sample preparation protocol used. All four GBS-positive samples identified by the standard culture method were detected by the two PCR assays. Conclusion: These assays provide promising tools for the rapid detection and identification of GBS.

Real time PCR assay for differentiation of Brucella abortus and Brucella melitensis

2017 ◽  
Author(s):  
Vinay Kumar ◽  
Sushila Maan ◽  
Aman Kumar ◽  
Kanisht Batra ◽  
Deepika Deepika ◽  
...  
Keyword(s):  
Real Time ◽  
Brucella Abortus ◽  
Real Time Pcr ◽  
Brucella Melitensis ◽  
Economic Losses ◽  
Conventional Pcr ◽  
Serological Tests ◽  
Detectable Difference ◽  
Pcr Assays ◽  
Respective Species

Brucellosis is one of the zoonotic diseases of major concern and can cause huge economic losses to livestock industry. Serological tests and bacterial isolation are considered as the gold standard assay for diagnosis of Brucella spp. but they are time-consuming, hazardous and lack specificity. To control and eradicate a disease, a confirmatory diagnostic method which is sensitive, quick and specific is the foremost requirement. Therefore in this study, we evaluated the performances of two newly designed TaqMan real-time PCR assays targeting the BruAB_0168 gene and BMEII0466 gene for Brucella abortus and Brucella melitensis (respectively). Both the assays were found to be highly specific in differentiation of respective species. Both the assays can detect as low as 0.02 fg of DNA and there was no detectable difference found in sensitivity of these two tests. R2 value and efficiency of these tests ranged from 0.992 - 0.998 and 100- 106%, respectively showing that these assays are highly efficient. Compared to conventional PCR assays these qPCR assays were 100 times higher sensitive. In conclusion, the present study showed that the developed real-time PCR assays are more sensitive, specific, have high reproducibility and repeatability and are faster than serological and conventional PCR methods for differentiation of Brucella abortus and Brucella melitensis.

scholarly journals Diagnosis of canine leptospirosis: evaluation of two PCR assays in comparison with the microagglutination test

2019 ◽  
Vol 39 (4) ◽  
pp. 255-262
Author(s):  
Paula L. Martin ◽  
Nestor O. Stanchi ◽  
Bibiana F. Brihuega ◽  
Estela Bonzo ◽  
Lucía Galli ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Whole Blood ◽  
Urine Samples ◽  
Clinical Samples ◽  
Conventional Pcr ◽  
Serum Samples ◽  
Presumptive Diagnosis ◽  
Microagglutination Test ◽  
Pcr Assays

ABSTRACT: Canine leptospirosis is definitely diagnosed by demonstrating seroconversion in paired serum samples from the acute and convalescent period by the microagglutination test (MAT). However, the application of a polymerase chain reaction (PCR) assay can provide earlier confirmation of suspected cases. The objective of this study was to evaluate two PCR assays used in diagnosis of human leptospirosis (lipL32 real-time PCR and rrs conventional PCR) in cultured microorganisms and experimentally contaminated samples (whole blood, serum, urine), and investigate their applicability in clinical samples from dogs with presumptive diagnosis of leptospirosis by using the MAT as a reference. The analytical sensitivity of the lipL32 real-time PCR was 1 genome equivalent per reaction, whereas that for the rrs conventional PCR was 10 genome equivalents per reaction. Both assays amplified the pathogenic strains but were negative when evaluating the DNA of other microorganisms that may be present in clinical samples. The lipL32 real-time PCR detected 100 bacteria/mL in whole blood samples, 1000 bacteria/mL in serum samples and 10 bacteria/mL in urine samples, whereas the rrs conventional PCR detected 1000 bacteria/mL in whole blood and serum samples and 100 bacteria/mL in urine samples. Seven out of the 51 samples from dogs with presumptive diagnosis of leptospirosis were considered as confirmed cases. ThelipL32 real-time PCR detected positive results in six of the seven confirmed cases, whereas the rrs conventional PCR detected four. The PCR assays evaluated proved to be useful diagnostic tools in the confirmation of canine leptospirosis when used together with the MAT.

Sign in / Sign up

Export Citation Format

Share Document