Comparison of laboratory methods for diagnosis of Trichomonas vaginalis

The paper presents data on comparison of microscopy, culture and molecular (PCR) methods for diagnosis of trichomoniasis, as well as on evaluation of the quality of culture media for isolation of trichomonads produced by Russian and international manufacturers. The highest sensitivity was shown for the molecular (real-time PCR) method. Microscopy of stained and wet smears demonstrated relatively high sensitivity. The culture media that are widely used in our country for Trichomonas vaginalis isolation need optimization.

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Development of qualitative methods of bifidobacterium bifidum in probiotic with Real-time PCR method

Heavy metals and arsenic concentrations in water, agricultural soil, and rice in Ngan Son district, Bac Kan province, Vietnam ◽

10.47866/2615-9252/vjfc.46 ◽

2018 ◽

Vol 1 (1) ◽

pp. 13-17

Author(s):

Trong Pham Nhu ◽

Long Le Thanh ◽

Trung Nguyen Thanh ◽

Yen Ta Thi ◽

Loan Pham Thi ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Dairy Products ◽

Food Additives ◽

Bifidobacterium Bifidum ◽

Pcr Assay ◽

Accurate Identification ◽

Powdered Milk ◽

Pcr Method

Bifidobacterium strains with probiotic effects have been widely used in dairy products, food additives and pharmaceuticals. Especially, Bifidobacterium bifidum (B. bifidum) is usually presented into food products such as functional food. However, it is difficult to detect, and quantify B. bifidum in a sample with a combination of different probiotics. In Vietnam, there is no official standard method to identify and quantify B. bifidum in the sample with the mix of probiotic species. To fulfil the requirements of a robust quality management, we have developed a quantitative real-time PCR assay based on groEL gene for accurate identification and quantification of Bifidobacterium bifidum. The developed assay allows an unambiguous speciesspecific detection. We built the real-time PCR method to detect and identify B. bifidum in functional and supplemented food with specific up to 100% and reproducibility (SR<0.25) suitable with Annex F AOAC: 2016. This real-time PCR method is rapidly and effectively than conventional method. It takes only 24 hours to detect and identify B. bifidum in compare with at least a period of 3-5 days for conventional methods. The low quantitative limit is 105 CFU/g/mL, which is consistent with probiotic and powdered milk products with a declared quality of more than 106 CFU/g/mL.

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A multiplex real-time PCR method for differentiation of beef, buffalo meat and pork

The Journal of Agriculture and Development ◽

10.52997/jad.8.01.2019 ◽

2019 ◽

Vol 18 (01) ◽

pp. 63-71

Author(s):

Tan M. Tran

Keyword(s):

Detection Limit ◽

Real Time ◽

Real Time Pcr ◽

High Sensitivity ◽

Specific Probe ◽

Food Authenticity ◽

Processed Meats ◽

Buffalo Meat ◽

Heat Treated ◽

Pcr Method

The objective of this study was to optimize a multiplex real-time PCR protocol for detection of DNA of beef, buffalo meat and pork, serving for food authenticity. The optimized concentrations were 200 nM primer and 100 nM specific probe for beef/buffalo meat, and 300 nM primer and 150 nM probe for pork. The amplification was performed using initial denaturation at 50oC for 2 min, 95oC for 2 min, followed by 45 cycles of denaturation at 95oC for 15 sec, and annealing and extension at 60oC for 40 sec. This protocol had high sensitivity and specificity. The detection limit of this method was found to be 0.1% in raw and heat-treated meat mix (80 – 121oC/15 min) or 0.005 ng DNA/reaction. The protocol of testing was applied for the commercial products both fresh and processed meats. The results demonstrated that 50% of raw beef sample (6/12) weren't found beef DNA. Eight of twelve beef sausage samples (66.67%) contained buffalo DNA. Beef DNA were found in all 12 samples of beef meatballs, but eight out of the 12 meatball samples were confirmed to have buffalo DNA (66.67%) and two out of the 12 meatball samples (16.67%) also contained porcine DNA.

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Identification of Etiologic Agents of the Pertussis-like Syndrome in Children by Real-time PCR Method

Prague Medical Report ◽

10.14712/23362936.2018.6 ◽

2018 ◽

Vol 119 (1) ◽

pp. 61-69 ◽

Cited By ~ 4

Author(s):

Shima Mahmoudi ◽

Maryam Banar ◽

Babak Pourakbari ◽

Hediyeh Sadat Alavi ◽

Hamid Eshaghi ◽

...

Keyword(s):

Influenza Virus ◽

Real Time ◽

Real Time Pcr ◽

Clinical Symptoms ◽

Cross Sectional ◽

Viral Pathogens ◽

Laboratory Methods ◽

Etiologic Agents ◽

Pcr Method ◽

Syncytial Virus

The aim of this study was to recognize the identity and frequency of etiologic agents of the pertussis-like syndrome in children < 2 years of age. A cross-sectional hospital-based study conducted from August 2014 to August 2015. All children < 2 years of age (n=100) who were suspected as pertussis infected were enrolled in this study and tested for Bordetella pertussis, adenovirus (Adv), respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and influenza virus A (INF-A) by real-time PCR technique. RSV was the most detected pathogen (20%), followed by B. pertussis (18%), Adv (16%), INF-A (11%), and hMPV (10%). Co-infection was observed in 8 patients (11%) and the combinations of RSV/INF-A (n=3, 4%), and AdV/B. pertussis (n=3, 4%) were more frequent. RSV, B. pertussis, and hMPV were more frequent pathogens among infants < 4 months of age. However, Adv and INF-A were more frequent pathogens among children > 6 months of age. In this study, RSV was the most frequent identified pathogen (n=20, 20%), followed by B. pertussis (n=18, 18%) and AdV (n=16, 16%). Pertussis was more frequent in spring (8%) and summer (6%). In addition, clinical symptoms of pertussis were the same as some viral pathogens, which can lead to misdiagnosis of infection. Therefore, diagnosis of pertussis should be established on the bases of both the clinical symptoms and the laboratory methods.

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

Anti-Infective Agents ◽

10.2174/2211352518999200629165108 ◽

2020 ◽

Vol 18 ◽

Author(s):

Pegah Shakib ◽

Mohammad Reza Zolfaghari

Keyword(s):

Streptococcus Pneumoniae ◽

Real Time ◽

Real Time Pcr ◽

False Negative ◽

Cross Sectional Study ◽

Laboratory Culture ◽

Cross Sectional ◽

Negative Results ◽

False Negative Results ◽

Pcr Method

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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Single-copy detection of somatic variants from solid and liquid biopsy

Scientific Reports ◽

10.1038/s41598-021-85545-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ana-Luisa Silva ◽

Paulina Klaudyna Powalowska ◽

Magdalena Stolarek ◽

Eleanor Ruth Gray ◽

Rebecca Natalie Palmer ◽

...

Keyword(s):

Real Time ◽

Single Molecule ◽

Real Time Pcr ◽

Clinical Decision Making ◽

High Sensitivity ◽

Clinical Decision ◽

Single Copy ◽

Turnaround Time ◽

Copy Detection ◽

Allele Specific

AbstractAccurate detection of somatic variants, against a background of wild-type molecules, is essential for clinical decision making in oncology. Existing approaches, such as allele-specific real-time PCR, are typically limited to a single target gene and lack sensitivity. Alternatively, next-generation sequencing methods suffer from slow turnaround time, high costs, and are complex to implement, typically limiting them to single-site use. Here, we report a method, which we term Allele-Specific PYrophosphorolysis Reaction (ASPYRE), for high sensitivity detection of panels of somatic variants. ASPYRE has a simple workflow and is compatible with standard molecular biology reagents and real-time PCR instruments. We show that ASPYRE has single molecule sensitivity and is tolerant of DNA extracted from plasma and formalin fixed paraffin embedded (FFPE) samples. We also demonstrate two multiplex panels, including one for detection of 47 EGFR variants. ASPYRE presents an effective and accessible method that simplifies highly sensitive and multiplexed detection of somatic variants.

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