Detection of Colorado Tick Fever viral RNA in acute human serum samples by a quantitative real-time RT-PCR assay

2007 ◽  
Vol 140 (1-2) ◽  
pp. 43-48 ◽  
Author(s):  
Amy J. Lambert ◽  
Olga Kosoy ◽  
Jason O. Velez ◽  
Brandy J. Russell ◽  
Robert S. Lanciotti
Keyword(s):  
Human Serum ◽  
Real Time ◽  
Viral Rna ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Serum Samples ◽  
Human Serum Samples

Comparison of seven commercial DNA extraction kits for the recovery of Brucella DNA from spiked human serum samples using real-time PCR

2007 ◽  
Vol 27 (2) ◽  
pp. 109-114 ◽  
Author(s):  
M. I. Queipo-Ortuño ◽  
F. Tena ◽  
J. D. Colmenero ◽  
P. Morata
Keyword(s):  
Human Serum ◽  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Serum Samples ◽  
Human Serum Samples

Detection of a novel bunyavirus associated with fever, thrombocytopenia and leukopenia syndrome in Henan Province, China, using real-time reverse transcription PCR

2013 ◽  
Vol 62 (7) ◽  
pp. 1060-1064 ◽  
Author(s):  
Xueyong Huang ◽  
Licheng Liu ◽  
Yanhua Du ◽  
Hongxia Ma ◽  
Yujiao Mu ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
High Sensitivity ◽  
Cross Reactivity ◽  
Henan Province ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Serum Samples ◽  
Reverse Transcription Pcr

A novel bunyavirus associated with fever, thrombocytopenia and leukopenia syndrome (FTLS) was discovered in Henan Province, China. Here, we report the development of an assay for this novel bunyavirus based on real-time reverse transcription PCR (RT-PCR). The assay exhibited high sensitivity and specificity without cross-reactivity towards 13 other viruses that cause similar symptoms. To evaluate the performance of this assay in detecting clinical samples, we analysed 261 serum samples from patients in Henan Province between 2007 and 2010. Of these samples, 91.95 % were bunyavirus positive. Compared with serological assays, the real-time PCR assay was much more sensitive in identifying infected patients 1 to 7 days after the onset of symptoms.

scholarly journals Clinical Evaluation of a New PCR Assay for Detection of Coxiella burnetii in Human Serum Samples

1998 ◽  
Vol 36 (1) ◽  
pp. 77-80 ◽  
Author(s):  
G. Q. Zhang ◽  
Sa V. Nguyen ◽  
H. To ◽  
M. Ogawa ◽  
A. Hotta ◽  
...  
Keyword(s):  
Human Serum ◽  
Coxiella Burnetii ◽  
Nested Pcr ◽  
Restriction Enzymes ◽  
Pcr Assay ◽  
Serum Samples ◽  
Gene Encoding ◽  
Human Serum Samples ◽  
Positive Serum ◽  
Pcr Method

A nested PCR method was developed for the detection ofCoxiella burnetii in human serum samples. Two pairs of oligonucleotide primers were designed to amplify a 438-bp fragment of the com1 gene encoding a 27-kDa outer membrane protein ofC. burnetii. The primers amplified the predicted fragments of 21 various strains of C. burnetii but did not react with DNA samples from other microorganisms. The 438-bp amplification products could be digested with restriction enzymes SspI and SalI. The utility of the nested PCR was evaluated by testing human serum samples. The com1 gene fragment was amplified from 135 (87%) of 155 indirect immunofluorescence test (IF)-positive serum samples and from 11 (11%) of 100 IF-negative serum samples. The nested PCR with primers targeted to the com1gene appeared to be a sensitive, specific, and useful method for the detection of C. burnetii in serum samples.

Real-time PCR for detection of Brucella spp. DNA in human serum samples

2005 ◽  
Vol 24 (12) ◽  
pp. 842-845 ◽  
Author(s):  
C. Debeaumont ◽  
P. A. Falconnet ◽  
M. Maurin
Keyword(s):  
Human Serum ◽  
Real Time ◽  
Real Time Pcr ◽  
Serum Samples ◽  
Human Serum Samples

scholarly journals A35 The first laboratory confirmation of chikungunya outbreak in Ethiopia

2019 ◽  
Vol 5 (Supplement_1) ◽  
Author(s):  
Mesfin Mengesha Tsegaye ◽  
Aadamu Tayachew ◽  
Desalegn Belay ◽  
Abebe Alemu ◽  
Berhane Beyene
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Laboratory Investigation ◽  
Emergency Preparedness ◽  
Zika Virus ◽  
Chikungunya Virus ◽  
Febrile Illness ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Serum Samples

Abstract Chikungunya is a viral disease (genus Alphavirus) which is transmitted to humans by infected mosquitoes—including Aedes aegypti and A. albopictus. An outbreak of febrile illness, suspected to have been caused by chikungunya, was reported in June 2016 from Dolloado district, Suuf Kebele, in the Somalia regional state of Ethiopia that borders the Mandera county of Kenya where a confirmed chikungunya outbreak was ongoing. Laboratory investigation was carried out to confirm if the outbreak in Ethiopia was caused by Chikungunya virus. Ten serum samples were collected from suspected patients visiting a health center in Suuf Kebel, who were then sent to the Nation laboratory in Ethiopian Public Health Institute. RNA was extracted from the serum samples using QIAgene RNA Mini kit, and PCR detection of dengue, chikungunya, and Zika virus nucleic acid was done using Trioplex Real-time RT-PCR Assay following the protocol from the Center for Disease Control (CDC). The Trioplex Real-time RT-PCR assay, for detection and differentiation of RNA from dengue, Chikungunya and Zika, was provided by CDC as part of the zika emergency preparedness effort. Of the nine samples tested, eight (88.88%) were found to be positive for chikungunya virus nucleic acid but negative for dengue and Zika virus nucleic acids. The median age of the affected sampled patients was 40 years, and males appear to be more affected (66.6% of sampled patients). The laboratory investigation confirmed that the outbreak was caused by chikungunya virus. Even though further molecular characterization of the positive isolates will provide more information as to the circulating genotypes and elucidate the origin of the outbreak virus, it is also possible to assume that the outbreak was an extension of the outbreak in neighboring countries in Kenya and, therefore, warrants that cross-border integration efforts to control chikungunya should be implemented by the concerned countries.

scholarly journals Single-step label-free nanowell immunoassay accurately quantifies serum stress hormones within minutes

2021 ◽  
Vol 7 (27) ◽  
pp. eabf4401
Author(s):  
S. Reza Mahmoodi ◽  
Pengfei Xie ◽  
Daniel P. Zachs ◽  
Erik J. Peterson ◽  
Rachel S. Graham ◽  
...  
Keyword(s):  
Human Serum ◽  
Real Time ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Label Free ◽  
Serum Samples ◽  
Electrode Gap ◽  
Buffer Solution ◽  
Human Serum Samples ◽  
Free Cortisol

A non-faradaic label-free cortisol sensing platform is presented using a nanowell array design, in which the two probe electrodes are integrated within the nanowell structure. Rapid and low volume (≤5 μl) sensing was realized through functionalizing nanoscale volume wells with antibodies and monitoring the real-time binding events. A 28-well plate biochip was built on a glass substrate by sequential deposition, patterning, and etching steps to create a stack nanowell array sensor with an electrode gap of 40 nm. Sensor response for cortisol concentrations between 1 and 15 μg/dl in buffer solution was recorded, and a limit of detection of 0.5 μg/dl was achieved. Last, 65 human serum samples were collected to compare the response from human serum samples with results from the standard enzyme-linked immunosorbent assay (ELISA). These results confirm that nanowell array sensors could be a promising platform for point-of-care testing, where real-time, laboratory-quality diagnostic results are essential.

Simultaneous Monitoring of Febuxostat and Uric Acid in Human Serum Samples Using the Direct Square-Wave Voltammetric Method

2019 ◽  
Vol 15 (6) ◽  
pp. 678-684
Author(s):  
Biljana Nigović ◽  
Jakov Vlak
Keyword(s):  
Uric Acid ◽  
Human Serum ◽  
Oxidase Inhibitor ◽  
Serum Samples ◽  
Boron Doped Diamond ◽  
Fast Analysis ◽  
Xanthine Oxidase Inhibitor ◽  
Voltammetric Method ◽  
Square Wave ◽  
Human Serum Samples

Background: High uric acid serum level, hyperuricemia, is now associated with many diseases such as gout, chronic kidney disease, hypertension, coronary artery disease and diabetes. Febuxostat is a novel selective xanthine oxidase inhibitor approved for the treatment of hyperuricemia. Objective: The aim of this study was to develop a first analytical method for the simultaneous determination of febuxostat and uric acid. Methods: An unmodified boron-doped diamond electrode provided concurrent quantitation of drug at low levels and uric acid, which has clinical significance in the diagnosis and therapy of hyperuricemia, at relatively high concentrations. The direct square-wave voltammetric method was applied to the analysis of both analytes in human serum samples. Results: Under the optimized conditions, the linear response of peak current on febuxostat concentration was achieved in the range from 7.5 × 10-7 to 3 × 10-5 M, while uric acid showed two linear ranges of 5 × 10-6 - 5 × 10-5 M and 5 × 10-5 - 2 × 10-4 M. The method was successfully utilised for quantification of both analytes in human serum samples. Good recoveries were obtained without interference from common inorganic cations and anions as well as glucose, dopamine, ascorbic and folic acids at concentrations expected in physiological conditions. Conclusion: The great benefits of developed method are fast analysis (only 7.5 s for run), low cost and simplicity of performance.

scholarly journals Investigations, actions and learning from an outbreak of SARS-CoV-2 infection among healthcare workers in the United Kingdom

2020 ◽  
pp. 175717742097679
Author(s):  
Kordo Saeed ◽  
Emanuela Pelosi ◽  
Nitin Mahobia ◽  
Nicola White ◽  
Christopher Labdon ◽  
...  
Keyword(s):  
Real Time ◽  
Healthcare Workers ◽  
Healthcare Facility ◽  
Rt Pcr ◽  
Serum Samples ◽  
The United Kingdom ◽  
Key Issues ◽  
Current Infection ◽  
Polymerase Chain ◽  
A Current

Background: We report an outbreak of SARS coronavirus-2 (SARS-CoV-2) infection among healthcare workers (HCW) in an NHS elective healthcare facility. Methodology: A narrative chronological account of events after declaring an outbreak of SARS-CoV-2 among HCWs. As part of the investigations, HCWs were offered testing during the outbreak. These were: (1) screening by real-time reverse transcriptase polymerase chain reaction (RT- PCR) to detect a current infection; and (2) serum samples to determine seroprevalence. Results: Over 180 HCWs were tested by real-time RT-PCR for SARS-CoV-2 infection. The rate of infection was 15.2% (23.7% for clinical or directly patient-facing HCWs vs. 4.8% in non-clinical non-patient-facing HCWs). Of the infected HCWs, 57% were asymptomatic. Seroprevalence (SARS-CoV-2 IgG) among HCWs was 13%. It was challenging to establish an exact source for the outbreak. The importance of education, training, social distancing and infection prevention practices were emphasised. Additionally, avoidance of unnecessary transfer of patients and minimising cross-site working for staff and early escalation were highlighted. Establishing mass and regular screening for HCWs are also crucial to enabling the best care for patients while maintaining the wellbeing of staff. Conclusion: To our knowledge, this is the first UK outbreak report among HCWs and we hope to have highlighted some key issues and learnings that can be considered by other NHS staff and HCWs globally when dealing with such a task in future.

Sign in / Sign up

Export Citation Format

Share Document