Development of Raw Material Identification Method of Changnan-jeot and Gaiyang-jeot Using Multiplex PCR and Real-Time PCR

2021 ◽  
Vol 36 (4) ◽  
pp. 289-297
Author(s):  
Seong Seok Choi ◽  
◽  
Yong Bae Seo ◽  
Jong-Oh Kim ◽  
Ji-Young Yang ◽  
...  
Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 152 ◽  
Author(s):  
Vivornpun Sanprasert ◽  
Ruthairat Kerdkaew ◽  
Siriporn Srirungruang ◽  
Sarit Charuchaibovorn ◽  
Kobpat Phadungsaksawasdi ◽  
...  

Soil-transmitted helminths (STHs) are the most common intestinal parasites infecting humans worldwide. STH infections are a major cause of morbidity and disability. Accurate diagnostic tools are pivotal for assessing the exact prevalence of parasitic infections. Microscopic examination and culture techniques have been used to observe the presence of eggs or larvae of parasites in stool samples, but they are time-consuming and have low sensitivity. Therefore, accurate, simple, and inexpensive diagnostic techniques are still required for simultaneous detection of STH infections. Although molecular-based techniques, such as real-time PCR and multiplex real-time PCR, have been developed, they are not suitable for routine diagnosis due to the requirement for expensive reagents and instruments. In this study, we established a conventional multiplex PCR for simultaneous rapid detection of Ascaris lumbricoides, Necator americanus, and Strongyloides stercoralis in stool samples. Our results show that the multiplex PCR could detect the DNA of STHs at a very low target gene concentrations (lower than 1 pg) with no cross-amplification. Multiplex PCR had five times higher sensitivity than the formalin–ethyl acetate concentration technique (FECT) in the detection of multiple infections, and two times higher for detection of S. stercoralis. However, multiplex PCR was comparable to FECT in the detection of A. lumbricoides and N. americanus. In conclusion, this method could be used as an alternative method for the detection of STHs, especially for S. stercoralis.


2004 ◽  
Vol 46 (5) ◽  
pp. 551-554 ◽  
Author(s):  
Rina Uzuka ◽  
Hisashi Kawashima ◽  
Daisuke Hasegawa ◽  
Hiroaki Ioi ◽  
Masahiro Amaha ◽  
...  

2017 ◽  
Vol 185 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Qingqing Wu ◽  
Shengnan Xiang ◽  
Wenjun Wang ◽  
Jinyan Zhao ◽  
Jinhua Xia ◽  
...  

Author(s):  
A. O. Sementsova ◽  
A. N. Shikov ◽  
V. A. Ternovoy ◽  
A. V. Vinokurova ◽  
N. E. Kostina ◽  
...  

Presented are the data on the development and approbation of the method of Marburg, Ebola, and Lassa viruses identification based on real-time multiplex PCR with hybridization-fluorescent detection. This method is meant for the differential diagnostics of hemorrhagic fevers caused by these viruses. Displayed are the results of determination of multiplex PCR analytical sensitivity and specific activity.


Author(s):  
Paviter Kaur ◽  
N. S. Sharma ◽  
A. K. Arora ◽  
Deepti .

The present study was carried out to evaluate the conventional and molecular techniques for diagnosis of bovine brucellosis. A total of four isolates of Brucella abortus obtained from 100 clinical samples of foetal stomach contents, vaginal mucus and uterine discharges were characterized biochemically. The isolates were confirmed as Brucella spp. by PCR using B4/B5 primer pair and as B. abortus by Bruce Ladder multiplex PCR. By Hinic Real-time PCR, all the four isolates were confirmed as Brucella spp. with Ct values between 14-16. With DNA extracted from 40 clinical samples of foetal stomach contents, vaginal mucus and uterine discharges, Real time PCR appeared most sensitive of the three molecular and conventional techniques as it detected maximum number of positive samples.


2019 ◽  
Vol 102 (5) ◽  
pp. 1552-1566
Author(s):  
Yange Yang ◽  
Mingchang Liu ◽  
Na Niu ◽  
Hongyue Wang ◽  
Bin Wang ◽  
...  

Abstract Background: Public interest is growing for small berries in recent years because they are very delicious, low in energy, and full of bioactive compounds with potential health benefits. Similar to other food products, adulteration of small berry fruit products poses economic and safety problems to consumers. Objective: To protect consumers and regulate the small berry fruit products market, it is necessary to establish a robust method for detecting the authenticity. Methods: In this study, TaqMan-based real-time PCR assay was established for species identification of cranberry, raspberry, and blueberry to ensure authenticity of commercial small berry food products (pulp, dried fruit, fruit juice, jam, and puree). Results: Absolute detection limit was 0.1 pg/μL DNA for raspberries, 1 pg/μL DNA for blueberries, and 10 pg/μL DNA for cranberries. Practical LOD was 0.1% (v/v) for fresh juice. For processed juice, practical LOD was 1% for blueberry and red raspberries, 0.1% for black and yellow raspberries, and 5% for cranberry. Conclusions: The method was shown to be functional and effective to detect the raw material composition of cranberry, raspberry, and blueberry for commercial products. Highlights: TaqMan probe-based real-time PCR methods were designed to identify three small berries (blueberry, raspberry, and cranberry) in berry products. Efficient DNA recovery methods and detection strategy were established to ensure correct and sensitive testing of fresh small berries exhibited a detection limit of 0.1 to 10 pg/μL. The practical minimum detection levels were 0.1 to 5% in fresh and processed juice, including pasteurization and HTHP.


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