Protocol for the Use of a Rapid Real-Time PCR Method for the Detection of HIV-1 Proviral DNA Using Double-Stranded Primer

Background: Conventional laboratory culture-based methods for diagnosis of Streptococcus pneumoniae are time-consuming and yield false negative results. Molecular methods including real-time (RT)-PCR rapid methods and conventional PCR due to higher sensitivity and accuracy have been replaced instead traditional culture assay. The aim of the current study was to evaluate lytA gene for detection of Streptococcus pneumoniae in the cerebrospinal fluid of human patients with meningitis using real-time PCR assay. Material and Methods: In this cross-sectional study, a total of 30 clinical specimens were collected from patients in a period from September to December 2018. In order to evaluate the presence of lytA gene, conventional and real-time PCR methods were used without culture. Results: From 30 sputum samples five (16.66%) isolates were identified as S. pneumoniae by lytA PCR and sequencing. Discussion: In this research, an accurate and rapid real-time PCR method was used, which is based on lytA gene for diagnosis of bacteria so that it can be diagnosed. Based on the sequencing results, the sensitivity for detection of lytA gene was 100% (5/5).

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Real-Time PCR Assay for Detection and Enumeration of Dekkera bruxellensis in Wine

Applied and Environmental Microbiology ◽

10.1128/aem.69.12.7430-7434.2003 ◽

2003 ◽

Vol 69 (12) ◽

pp. 7430-7434 ◽

Cited By ~ 77

Author(s):

Trevor G. Phister ◽

David A. Mills

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Pcr Primers ◽

Rrna Gene ◽

Dekkera Bruxellensis ◽

Pcr Assay ◽

Specific Pcr ◽

Spoilage Yeast ◽

Pcr Method ◽

26S Rrna

ABSTRACT Traditional methods to detect the spoilage yeast Dekkera bruxellensis from wine involve lengthy enrichments. To overcome this difficulty, we developed a quantitative real-time PCR method to directly detect and enumerate D. bruxellensis in wine. Specific PCR primers to D. bruxellensis were designed to the 26S rRNA gene, and nontarget yeast and bacteria common to the winery environment were not amplified. The assay was linear over a range of cell concentrations (6 log units) and could detect as little as 1 cell per ml in wine. The addition of large amounts of nontarget yeasts did not impact the efficiency of the assay. This method will be helpful to identify possible routes of D. bruxellensis infection in winery environments. Moreover, the time involved in performing the assay (3 h) should enable winemakers to more quickly make wine processing decisions in order to reduce the threat of spoilage by D. bruxellensis.

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Development and application of a novel reverse transcription real-time PCR method for miR-499 quantification

Clinical Biochemistry ◽

10.1016/j.clinbiochem.2013.06.024 ◽

2013 ◽

Vol 46 (15) ◽

pp. 1566-1571 ◽

Cited By ~ 10

Author(s):

Weidong Zheng ◽

Yuwei Di ◽

Yinghong Liu ◽

Ge Huang ◽

Youwei Zheng ◽

...

Keyword(s):

Real Time ◽

Reverse Transcription ◽

Real Time Pcr ◽

Pcr Method

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Protocol for the Use of a Rapid Real-Time PCR Method for the Detection of HIV-1 Proviral DNA Using Double-Stranded Primer

A rapid real-time PCR assay for the detection of HIV-1 proviral DNA using double-stranded primer

WITHDRAWN: Rapid, point-of-care extraction of HIV-1 proviral DNA from whole blood for detection by real-time PCR

Quantitation of HIV-1 group M proviral DNA using TaqMan MGB real-time PCR

Evaluating the HIV-1 Proviral DNA Detection by Use of Real Time PCR from Blood Samples and Dried Blood Spots

A simple and rapid DNA extraction method from whole blood for highly sensitive detection and quantitation of HIV-1 proviral DNA by real-time PCR

Comparison of real-time PCR methods for measurement of HIV-1 proviral DNA

Human immunodeficiency virus type 1 (HIV-1) proviral DNA load in purified CD4+ cells by LightCycler® Real-time PCR

Detection of Streptococcus pneumoniae in Sputum Samples by Real- Time PCR.

Real-Time PCR Assay for Detection and Enumeration of Dekkera bruxellensis in Wine

Development and application of a novel reverse transcription real-time PCR method for miR-499 quantification

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