scholarly journals Microarray analysis and real-time PCR assay developed to find biomarkers for mercury-contaminated soil

2016 ◽  
Vol 5 (6) ◽  
pp. 1539-1547 ◽  
Author(s):  
Jing Hou ◽  
Xinhui Liu ◽  
Baoshan Cui ◽  
Junhong Bai ◽  
Xiangke Wang
Keyword(s):  
Adverse Effects ◽  
Human Health ◽  
Real Time ◽  
Microarray Analysis ◽  
Food Chain ◽  
Contaminated Soil ◽  
Real Time Pcr ◽  
Agricultural Soil ◽  
Pcr Assay ◽  
Soil P

The evaluation of mercury (Hg) toxicity in agricultural soil is of great concern because its bioavailability and bioaccumulation in organisms through the food chain can have adverse effects on human health.

Molecular Detection of the Three Major Pathogenic Vibrio Species from Seafood Products and Sediments in Tunisia Using Real-Time PCR

2016 ◽  
Vol 79 (12) ◽  
pp. 2086-2094 ◽  
Author(s):  
MORSI GDOURA ◽  
HANEN SELLAMI ◽  
HANEN NASFI ◽  
RAHMA TRABELSI ◽  
SABEUR MANSOUR ◽  
...  
Keyword(s):  
Human Health ◽  
Real Time ◽  
Real Time Pcr ◽  
Ruditapes Decussatus ◽  
Conventional Pcr ◽  
Pcr Assay ◽  
Isolation And Characterization ◽  
Seafood Products ◽  
Pcr Method ◽  
Vibrio Spp

ABSTRACT Vibrio spp. have emerged as a serious threat to human health worldwide. V. parahaemolyticus, V. cholerae, and V. vulnificus pose a considerable public health risk in Tunisia because they cause sporadic and epidemic foodborne infections associated with the consumption of raw or undercooked contaminated seafood. More recently, toxR-positive V. alginolyticus was also reported to be a potential source of contaminated seafood. A total of 247 samples, including 113 fishes (Labrus viridis, Penaeus kerathurus, Diplodus annularis, Diplodus sparaillon, Scorparna porcus, Sarpa salpa, Dentex dentex, Scorparna scrofa, Sardinella aurita, Trachurus trachurus, Synodus saurus, Pagellus erythrinus, and Metapenaeus monoceros), 83 clams (Ruditapes decussatus species), 30 seawater samples, and 21 sediment samples were analyzed using traditional culture methods (ISO/TS 21872-1; International Organization for Standardization 2007) and a conventional PCR method for Vibrio spp. identification. A rapid, sensitive, and highly reproducible real-time PCR assay was developed to detect the three major Vibrio spp. pathogenic for humans in Tunisian seafood products and sediments. A conventional culture method found 102 (41.3%) of 247 analyzed samples positive for Vibrio spp.; a conventional PCR method found 126 (51%) of the 247 samples positive. Real-time PCR assay found 126 (51.1%) samples positive; V. alginolyticus toxR was the most common, found in 99 (78.57%) of samples, followed by V. parahaemolyticus in 26 (20.63%) and V. cholerae in 1 (0.7%). All culture-positive samples were PCR positive. However, 24 samples that were positive by conventional PCR and real-time PCR were culture negative. Our findings indicate that retail seafood is commonly contaminated with Vibrio spp. and presents a potential risk to human health in Tunisia. These data also indicate that real-time PCR can provide sensitive species-specific detection of Vibrio spp. in seafood without prior isolation and characterization of the bacteria by traditional microbiological methods.

scholarly journals A misleading false-negative result of Pneumocystis real-time PCR assay due to a rare punctual mutation: A French multicenter study

2016 ◽  
Vol 55 (2) ◽  
pp. 180-184 ◽  
Author(s):  
Solène Le Gal ◽  
Florence Robert-Gangneux ◽  
Yann Pépino ◽  
Sorya Belaz ◽  
Céline Damiani ◽  
...  
Keyword(s):  
Real Time ◽  
Multicenter Study ◽  
Real Time Pcr ◽  
False Negative ◽  
False Negative Result ◽  
Pcr Assay

scholarly journals Differing Effects of Standard and Harsh Nucleic Acid Extraction Procedures on Diagnostic Helminth Real-Time PCRs Applied to Human Stool Samples

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 188
Author(s):  
Tanja Hoffmann ◽  
Andreas Hahn ◽  
Jaco J. Verweij ◽  
Gérard Leboulle ◽  
Olfert Landt ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Acid Extraction ◽  
Nucleic Acid Extraction ◽  
Pcr Assay ◽  
Bead Beating ◽  
Stool Samples ◽  
Pcr Assays ◽  
Human Stool

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.

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