Development and Evaluation of a Real-Time PCR Assay for Detection and Quantification of Blastocystis Parasites in Human Stool Samples: Prospective Study of Patients with Hematological Malignancies

This study aimed to assess standard and harsher nucleic acid extraction schemes for diagnostic helminth real-time PCR approaches from stool samples. A standard procedure for nucleic acid extraction from stool and a procedure including bead-beating as well as proteinase K digestion were compared with group-, genus-, and species-specific real-time PCR assays targeting helminths and nonhelminth pathogens in human stool samples. From 25 different in-house and commercial helminth real-time PCR assays applied to 77 stool samples comprising 67 historic samples and 10 external quality assessment scheme samples positively tested for helminths, higher numbers of positive test results were observed after bead-beating-based nucleic acid extraction for 5/25 (20%) real-time PCR assays irrespective of specificity issues. Lower cycle threshold values were observed for one real-time PCR assay after the standard extraction scheme, and for four assays after the bead-beating-based scheme. Agreement between real-time PCR results after both nucleic acid extraction strategies according to Cohen’s kappa ranged from poor to almost perfect for the different assays. Varying agreement was observed in eight nonhelminth real-time PCR assays applied to 67 historic stool samples. The study indicates highly variable effects of harsh nucleic acid extraction approaches depending on the real-time PCR assay used.

Download Full-text

Development of a TaqMan based real-time PCR assay for detection of Clonorchis sinensis DNA in human stool samples and fishes

Parasitology International ◽

10.1016/j.parint.2011.06.010 ◽

2012 ◽

Vol 61 (1) ◽

pp. 183-186 ◽

Cited By ~ 23

Author(s):

Xian-Quan Cai ◽

Hai-Qiong Yu ◽

Jian-Shan Bai ◽

Jian-Dong Tang ◽

Xu-Chu Hu ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Clonorchis Sinensis ◽

Pcr Assay ◽

Stool Samples ◽

Human Stool

Download Full-text

Detection of Peanut Allergen Traces with a Real Time PCR Assay - The Challenge to Protect Food-Allergic Consumers

Journal of Food Research ◽

10.5539/jfr.v7n1p32 ◽

2017 ◽

Vol 7 (1) ◽

pp. 32 ◽

Cited By ~ 2

Author(s):

Dimitra Houhoula ◽

Stamatios Koussissis ◽

Vladimiros Lougovois ◽

John Tsaknis ◽

Dimitra Kassavita ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Rapid Detection ◽

Food Products ◽

Molecular Techniques ◽

Food Allergens ◽

Pcr Assay ◽

Peanut Allergen ◽

Pcr Method ◽

Detection And Quantification

The aim of the present study was the implementation of molecular techniques in the detection and quantification of allergic substances of peanut in various kinds of food products, e.g., breakfast cereals, chocolates and biscuits that are frequently related to allergies. In some cases, the presence of peanuts can be due to contamination during production and are not declared on the label. A total of 152 samples were collected from supermarkets and were analysed by a Real Time PCR method. The results indicated that 125 samples (83,3%) were found positive in peanut traces but the most important finding is that from the 84 samples that had no allergen declaration for peanuts, 48 (57,1%) of them were found positive. In conclusion, Real Time PCR can be a very important tool for the rapid detection and quantification of food allergens.

Download Full-text