scholarly journals Comparison between LightCycler Real-Time Polymerase Chain Reaction (PCR) Assay with Serum and PCR-Enzyme-Linked Immunosorbent Assay with Whole Blood Samples for the Diagnosis of Human Brucellosis

2005 ◽  
Vol 40 (2) ◽  
pp. 260-264 ◽  
Author(s):  
M. I. Queipo-Ortuno ◽  
J. D. Colmenero ◽  
G. Baeza ◽  
P. Morata
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Whole Blood ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Brucellosis ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Blood Samples ◽  
Polymerase Chain ◽  
Whole Blood Samples

scholarly journals Detection of Genetically Modified Corn (Bt176) in Spiked Cow Blood Samples by Polymerase Chain Reaction and Immunoassay Methods

2005 ◽  
Vol 88 (2) ◽  
pp. 654-664 ◽  
Author(s):  
Laetitia Petit ◽  
Fabienne Baraige ◽  
Yves Bertheau ◽  
Philippe Brunschwig ◽  
Annick Diolez ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Whole Blood ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Cry1ab Protein ◽  
Blood Samples ◽  
Polymerase Chain ◽  
Whole Blood Samples

Abstract The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35S promoter and Bt176-specific junction sequence) was determined by both the polymerase chain reaction–enzyme-linked immunosorbent assay (PCR–ELISA) and the 5′-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (αs1-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/mL in 10 mL whole blood samples. The sandwich ELISA kit used detected down to 1 ng transgenic Cry1Ab protein/mL spiked whole blood.

Screening for Aspergillus spp. using polymerase chain reaction of whole blood samples from patients with haematological malignancies

2001 ◽  
Vol 113 (1) ◽  
pp. 180-184 ◽  
Author(s):  
Cornelia Lass-Flörl ◽  
Johannes Aigner ◽  
Eberhard Gunsilius ◽  
Andreas Petzer ◽  
David Nachbaur ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Whole Blood ◽  
Haematological Malignancies ◽  
Chain Reaction ◽  
Blood Samples ◽  
Polymerase Chain ◽  
Whole Blood Samples

scholarly journals Direct Amplification of B1 gene of Toxoplasma gondii DNA using Nested Polymerase Chain Reaction Following Microwave Treatment for Whole Blood Samples

2015 ◽  
Vol 39 (1) ◽  
pp. 23-27
Author(s):  
Balkes Fadel Hade
Keyword(s):  
Polymerase Chain Reaction ◽  
Whole Blood ◽  
Microwave Treatment ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
B1 Gene ◽  
Polymerase Chain ◽  
Whole Blood Samples ◽  
Commercial Kit

     Infection with Toxoplasma gondii is a cause of fetal death since T. gondii can be transmitted to the fetus through the placenta (transplacental) from an infected mother or at vaginal delivery. Blood obtained from women and sheep to confirm their infection with toxoplasmosis by using Enzyme Linked Immunosorbant Assay test (ELISA) to ditective positive specific anti-Toxoplasma (IgM, IgG and IgM, or IgG) antibodies. This study used two methodes to extract DNA (the first one was a standard extraction commercial method (CM-PCR) of genomic DNA using a commercial kit (Promega, USA), and the second one was the direct heat DNA extraction using microwave oven (MW-PCR) for whole blood samples obtained from infected women and sheep. Then nested Polymerase Chain Reaction (n PCR) were used to amplify Toxoplasma B1 gene to detect T. gondii DNA in whole blood samples. The results indecated using of microwave treatment instead of commercial kit to extract DNA is low cost and short time,and complement serology for clinical studies and   diagnostic purposes of toxoplasmosis.                                                                     

scholarly journals RH genotypes among Malaysian blood donors

2014 ◽  
Vol 8 (4) ◽  
pp. 499-504 ◽  
Author(s):  
Rozi Hanisa Musa ◽  
Afifah Hassan ◽  
Yasmin Ayob ◽  
Narazah Mohd Yusoff
Keyword(s):  
Polymerase Chain Reaction ◽  
Ethnic Groups ◽  
Whole Blood ◽  
Molecular Basis ◽  
Blood Donors ◽  
Limited Information ◽  
Chain Reaction ◽  
Blood Samples ◽  
Polymerase Chain ◽  
Whole Blood Samples

AbstractBackground: RH genotyping studies have been conducted mainly in people of Caucasian and African descent. There is limited information regarding the molecular basis for RH genotypes in Malaysia.Objectives: To investigate the prevalence and characteristics of RHCE genotypes among different ethnic groups in Malaysia.Methods: A total of 1014 whole blood samples were obtained from donors from 4 different ethnic groups (360 Malays, 434 Chinese, 164 Indians, and 56 others). All samples were phenotyped for C, c, D, E, and e using standard serologic methods and genotyped using polymerase chain reaction (PCR)-based analysis.Results: In the blood samples that we analyzed, the distribution of RH genotype antigens was significantly different among the various ethnic groups. Our findings showed that CCDee is the most common in Malaysian blood donors; 18.4% (187/1014) compared with other genotypes. The ccDEE genotype is more prevalent in the Chinese: 65.6% (82/125), and the ccee genotype is more prevalent in Indians: 47.1% (65/138). There were discrepancies between phenotypes and genotypes. There were 17 (1.7%) discrepancies in RH C/c genotyping results and of these 47% (8/17) occurred in Malays. Discrepancies in RH E/e results occurred in 3 samples (0.3%).Conclusions: Our study provides a database for the distribution of RH genotypes of donors from the major ethnic groups in Malaysia. Methods used in this study are useful for comparing the phenotypes and genotypes. Further investigation should be conducted to study the causes of these discrepancies using other molecular based techniques.

scholarly journals Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Qrt Pcr ◽  
Highly Sensitive ◽  
One Step ◽  
Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

scholarly journals Comparison of multiplex real-time polymerase chain reaction with serological tests and culture for diagnosing human brucellosis

2019 ◽  
Vol 12 (3) ◽  
pp. 337-342 ◽  
Author(s):  
Tuba Dal ◽  
Soner Sertan Kara ◽  
Aytekin Cikman ◽  
Cigdem Eda Balkan ◽  
Ziya Cibali Acıkgoz ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Human Brucellosis ◽  
Chain Reaction ◽  
Serological Tests ◽  
Polymerase Chain
Sign in / Sign up

Export Citation Format

Share Document