scholarly journals Highly sensitive and multiplexed analysis of CpG methylation at single-base resolution with ligation-based exponential amplification

2015 ◽  
Vol 6 (3) ◽  
pp. 1866-1872 ◽  
Author(s):  
Fengxia Su ◽  
Limei Wang ◽  
Yueying Sun ◽  
Chenghui Liu ◽  
Xinrui Duan ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Cpg Methylation ◽  
Chain Reaction ◽  
Single Base ◽  
Ligase Chain Reaction ◽  
Highly Sensitive ◽  
Single Base Resolution ◽  
Multiplexed Analysis

Multiple CpG methylation can be accurately detected in one-tube ligase chain reaction (LCR) amplification with high sensitivity and specificity.

Highly sensitive detection of CpG methylation in genomic DNA by AuNP-based colorimetric assay with ligase chain reaction

2015 ◽  
Vol 51 (16) ◽  
pp. 3371-3374 ◽  
Author(s):  
Fengxia Su ◽  
Limei Wang ◽  
Yueying Sun ◽  
Chenghui Liu ◽  
Xinrui Duan ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Genomic Dna ◽  
Colorimetric Assay ◽  
Colorimetric Detection ◽  
Cpg Methylation ◽  
Chain Reaction ◽  
Selective Method ◽  
Ligase Chain Reaction ◽  
Highly Sensitive ◽  
Highly Sensitive Detection

Using LCR amplification and AuNP-based colorimetric detection, a highly sensitive and selective method for detection of DNA methylation has been developed.

scholarly journals Gene specific-loci quantitative and single-base resolution analysis of 5-formylcytosine by compound-mediated polymerase chain reaction

2018 ◽  
Vol 9 (15) ◽  
pp. 3723-3728 ◽  
Author(s):  
Yafen Wang ◽  
Chaoxing Liu ◽  
Xiong Zhang ◽  
Wei Yang ◽  
Fan Wu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Demethylation ◽  
Chain Reaction ◽  
Single Base ◽  
Active Dna Demethylation ◽  
Key Players ◽  
Resolution Analysis ◽  
Single Base Resolution ◽  
Polymerase Chain

5-Formylcytosine (5fC) is known as one of the key players in the process of active DNA demethylation and displays essential epigenetic functions in mammals.

Polymerase chain reaction for the amplification of the 121-bp repetitive sequence ofSchistosoma mansoni: a highly sensitive potential diagnostic tool for areas of low endemicity

2014 ◽  
Vol 89 (6) ◽  
pp. 769-773 ◽  
Author(s):  
E. Ferrer ◽  
F. Pérez ◽  
I. Bello ◽  
A. Bolívar ◽  
M. Lares ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sensitivity And Specificity ◽  
Repetitive Sequence ◽  
Annealing Temperature ◽  
Immunological Methods ◽  
Molecular Technique ◽  
Analytical Sensitivity ◽  
Chain Reaction ◽  
Highly Sensitive ◽  
Polymerase Chain

AbstractSchistosomiasis is a disease caused by parasitic flatworms of the genusSchistosoma, whose diagnosis has limitations, such as the low sensitivity and specificity of parasitological and immunological methods, respectively. In the present study an alternative molecular technique requiring previous standardization was carried out using the polymerase chain reaction (PCR) for the amplification of a 121-bp highly repetitive sequence forSchistosoma mansoni.DNA was extracted from eggs ofS. mansoniby salting out. Different conditions were standardized for the PCR technique, including the concentration of reagents and the DNA template, annealing temperature and number of cycles, followed by the determination of the analytical sensitivity and specificity of the technique. Furthermore, the standardized PCR technique was employed in DNA extracted, using Chelex®100, from samples of sera of patients with an immunodiagnosis of schistosomiasis. The optimal conditions for the PCR were 2.5 mmMgCl2, 150 mmdeoxynucleoside triphosphates (dNTPs), 0.4 μmprimers, 0.75 U DNA polymerase, using 35 cycles and an annealing temperature of 63°C. The analytical sensitivity of the PCR was 10 attograms of DNA and the specificity was 100%. The DNA sequence was successfully detected in the sera of two patients, demonstrating schistosomiasis transmission, although low, in the community studied. The standardized PCR technique, using smaller amounts of reagents than in the original protocol, is highly sensitive and specific for the detection of DNA fromS. mansoniand could be an important tool for diagnosis in areas of low endemicity.

scholarly journals POLYMERASE CHAIN REACTION AS A DIAGNOSTIC TOOL FOR SIX SEXUALLY TRANSMITTED INFECTIONS - PRELIMINARY RESULTS

2015 ◽  
Vol 88 (1) ◽  
pp. 33-37
Author(s):  
Alecsandra Iulia Grad ◽  
Mihaela Laura Vica ◽  
Horea Vladi Matei ◽  
Doru Lucian Grad ◽  
Ioan Coman ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chlamydia Trachomatis ◽  
Neisseria Gonorrhoeae ◽  
Sexually Transmitted Infections ◽  
Sensitivity And Specificity ◽  
Diagnostic Tool ◽  
High Sensitivity ◽  
Chain Reaction ◽  
Sexually Transmitted ◽  
Polymerase Chain

Background and aim. Sexually transmitted infections are a very frequent and under-diagnosed cause of illness worldwide. A high number of detection methods and a large range of specimens in which sexually transmitted infections can be determined are available at the moment. Polymerase chain reaction performed on first void urine offers the advantage of being non-invasive, self-collectable and has high sensitivity and specificity. We looked to determine the frequency of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma hominis, Mycoplasma genitalium and Ureaplasma urealyticum in symptomatic and asymptomatic patients.Methods. Six sexually transmitted infections were determined in the first void urine of 15 symptomatic and asymptomatic patients by polymerase chain reaction. We used “Epicenter MasterPure™ Complete DNA and RNA Purification Kit” for the DNA purification and “Seeplex® STD6 ACE Detection” for the DNA amplification. The results were examined in UV light.Results. A number of 5 patients had positive results for Chlamydia trachomatis or Neisseria gonorrhoeae. Sexually transmitted infections are more frequent in men between 27 and 40 years old.Conclusions. Polymerase chain reaction is a good diagnostic tool for sexually transmitted infections because it has a high sensitivity and specificity. Chlamydia trachomatis is the most frequent sexually transmitted infection, followed by Neisseria gonorrhoeae.

scholarly journals LOOP-MEDIATED ISOTHERMAL AMPLIFICATION FOR DETECTION OF C. BURNETII IN BULGARIA

2021 ◽  
Vol 19 (2) ◽  
pp. 147-151
Author(s):  
M. Kunchev ◽  
V. Belcheva ◽  
E. Grigorov
Keyword(s):  
Polymerase Chain Reaction ◽  
Sensitivity And Specificity ◽  
Q Fever ◽  
High Sensitivity ◽  
Isothermal Amplification ◽  
Chain Reaction ◽  
Retrospective Diagnosis ◽  
Loop Mediated Isothermal Amplification ◽  
The World ◽  
Polymerase Chain

Q fever, which is caused by Coxiella burnetii, a small, pleomorphic intracellular bacterium, is the most widespread zoonosis in the world. The chronic form of the disease can lead to disability and death. Rapid diagnosis of Q fever is needed in order that effective treatment can be initiated. The conventional retrospective diagnosis of Q fever, based on serology, is useless for the treatment of afflicted patients. Thus, molecular methods have been created to close the diagnostic gap between the onset of the disease and the presence of specific antibodies in serum. A polymerase chain reaction is a suitable and reliable method with high sensitivity and specificity, but it requires expensive equipment and post-amplification protocol. Loop-mediated isothermal amplification (LAMP) is an isothermal technique, conducted at constant temperature that can amplify a negligible amount of DNA to more than 109 copies within one hour, using special primers and polymerase. We have tested the sensitivity and specificity of LAMP in the detection of C. burnetii. The mean positive rate of LAMP and polymerase chain reaction in patients was 100% and 74%, respectively. LAMP reacted negatively with non-C. burnetii pathogens and non-infected blood samples. We conclude that LAMP is a sensitive and specific technique for the detection of C. burnetii and has advantages over serological methods and PCR that make it attractive for diagnosing Q fever in countries around the world.

Highly sensitive DNA methylation analysis at CpG resolution by surface-enhanced Raman scattering via ligase chain reaction

2015 ◽  
Vol 51 (54) ◽  
pp. 10953-10956 ◽  
Author(s):  
Yuling Wang ◽  
Eugene J. H. Wee ◽  
Matt Trau
Keyword(s):  
Dna Methylation ◽  
Raman Scattering ◽  
Methylation Analysis ◽  
Chain Reaction ◽  
Ligase Chain Reaction ◽  
Highly Sensitive ◽  
Surface Enhanced ◽  
Surface Enhanced Raman ◽  
Enhanced Raman Scattering ◽  
Dna Methylation Analysis

Highly sensitive DNA methylation analysis at CpG resolution is demonstrated by employing SERS nanotags via ligase chain reaction (LCR) and validated with sequencing.

One-step detection of microRNA with high sensitivity and specificity via target-triggered loop-mediated isothermal amplification (TT-LAMP)

2017 ◽  
Vol 53 (80) ◽  
pp. 11040-11043 ◽  
Author(s):  
Yuanyuan Sun ◽  
Hui Tian ◽  
Chenghui Liu ◽  
Yueying Sun ◽  
Zhengping Li
Keyword(s):  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Isothermal Amplification ◽  
Sensitive Detection ◽  
Step Detection ◽  
Loop Mediated Isothermal Amplification ◽  
Highly Sensitive ◽  
One Step ◽  
Highly Sensitive Detection

A target-triggered loop-mediated isothermal amplification (TT-LAMP) mechanism is developed for simple one-step but highly sensitive detection of microRNAs.

scholarly journals Copro-PCR in the detection and confirmation of Toxoplasma gondii oocysts in feces of stray and domiciled cats

2021 ◽  
Vol 30 (2) ◽  
Author(s):  
Jaqueline Ataíde Silva Lima da Igreja ◽  
Hanstter Hallison Alves Rezende ◽  
Jade de Oliveira Melo ◽  
João Luís Garcia ◽  
Felippe Danyel Cardoso Martins ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Toxoplasma Gondii ◽  
Genetic Material ◽  
High Sensitivity ◽  
Molecular Techniques ◽  
Molecular Methods ◽  
Chain Reaction ◽  
Highly Sensitive ◽  
Polymerase Chain ◽  
Fecal Tests

Abstract Molecular methods such as Copro-PCR stand out in the diagnosis of T. gondii, because they are highly sensitive and specific, and can distinguish T. gondii from other morphologically similar coccids. The purpose was the detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction in 149 fecal samples from stray and domiciled cats, using three distinct markers (B5-B6, 18S and 529bp RE). Oocysts of T. gondii/H. hammondi were detected in 15.4% by parasitology fecal tests (PFT), and 4% of these oocysts were positively identified as T. gondii by Copro-PCR. The presence of T. gondii genetic material was detected in 16.1%, but 12% of the samples that tested positive by Copro-PCR were negative in PFT. Samples with discordant results were subjected to a new Copro-PCR with 18S marker and a 529, and of the 17 samples, 9 contained T. gondii genetic material. A comparison of the PFT and the molecular methods showed the latter was more sensitive, since it detected 22.1% while the PFT detected 15.4%. Demonstrating the high sensitivity and specificity of the Copro-PCR, particularly with the association of primers (k=0.809), but also confirms the importance of using molecular techniques in laboratories, since Copro-PCR was able to detect samples considered negative by PFT.

Molybdenum disulfide nanomaterial sensor-based detection of breast cancer microRNA

2020 ◽  
Vol 10 (6) ◽  
pp. 915-921 ◽  
Author(s):  
Runling Zhang ◽  
ChengJing Xia ◽  
Xiaohe Zhou ◽  
Yanni Guo ◽  
Weijie Li
Keyword(s):  
Breast Cancer ◽  
Molybdenum Disulfide ◽  
Detection Method ◽  
High Sensitivity ◽  
High Specificity ◽  
Fluorescence Sensor ◽  
Dna Probe ◽  
Single Base ◽  
Highly Sensitive ◽  
Before And After

In this study, we sought to establish a rapid, simple, highly sensitive, and highly specific fluorescence sensor for the detection of breast cancer-related microRNA (miRNA) based on the novel nanomaterial known as molybdenum disulfide (MoS2). MoS2, a novel nanomaterial, was added to a fluorescently-labeled DNA probe system to construct a fluorescence sensor. Furthermore, non-complementary miRNAs, single-base mismatched miRNAs, and different concentrations of complementary miRNAs were hybridized with the fluorescently-labeled DNA probe. Then, miRNAs were detected by measuring changes in the fluorescence signals before and after hybridization. The detection was completed within 40 min. The detection limit of complementary miRNA reached pmol/L levels while distinguishing between complementary, non-complementary, and single-base mismatched miRNAs. The MoS2 fluorescence sensor demonstrated rapid analysis, high sensitivity, and high specificity. Additionally, this established detection method has the potential for application in the rapid detection of breast cancer-related miRNA as a promising cancer detection method.

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