Universal primers for species authentication of animal foodstuff in a single polymerase chain reaction

2012 ◽  
Vol 93 (2) ◽  
pp. 354-361 ◽  
Author(s):  
Jose L Horreo ◽  
Alba Ardura ◽  
Ivan G Pola ◽  
Jose L Martinez ◽  
Eva Garcia-Vazquez
Keyword(s):  
Polymerase Chain Reaction ◽  
Universal Primers ◽  
Chain Reaction ◽  
Single Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Species Authentication

Streptococcus pneumoniae Serotyping by a Single Polymerase Chain Reaction–Based Multiplex Assay

2018 ◽  
Vol 26 (2) ◽  
pp. 75-79
Author(s):  
Mohammad Shokri Moghadam ◽  
Malihe Talebi ◽  
Faramarz Masjedian ◽  
Gholamreza Irajian ◽  
Mohammad Reza Pourshafie
Keyword(s):  
Polymerase Chain Reaction ◽  
Streptococcus Pneumoniae ◽  
Multiplex Assay ◽  
Chain Reaction ◽  
Single Polymerase Chain Reaction ◽  
Polymerase Chain

scholarly journals Simplified Method of Detection of Dialister invisus and Olsenella uli in Oral Cavity Samples by Polymerase Chain Reaction

2017 ◽  
Vol 8 (1-2) ◽  
pp. 47-52 ◽  
Author(s):  
Manohar S. Kugaji ◽  
Kishore G. Bhat ◽  
Vinayak M. Joshi ◽  
Madhu Pujar ◽  
Pratik T. Mavani
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacterial Species ◽  
Universal Primers ◽  
Sample Collection ◽  
Chain Reaction ◽  
Infection Group ◽  
Endodontic Infection ◽  
Predominant Component ◽  
Endodontic Infections ◽  
Polymerase Chain

Aims and Objectives: The oral microbial flora is highly complex and diverse with obligate anaerobic bacteria as the predominant component. Most of these are not yet cultivated/difficult to cultivate due to technical limitations. In this study, we aim to detect two novel oral bacterial species Dialister invisus and Olsenella uli by simplified and economical procedure of polymerase chain reaction (PCR) and study their association with primary and persistent endodontic infections. Material and Methods: The study involved 60 patients that included 30 patients of primary endodontic infections and 30 with persistent endodontic infections. The sample collection from the root canal was performed by universally accepted protocol by using sterile paper points. The deoxyribonucleic acid (DNA) extraction was done, followed by PCR with species specific primers. We made several changes to the protocol mentioned by original authors. We adopted a one-step protocol for amplification of bacterial DNA, omitting the 16SrDNA amplification step with universal primers. Results: It was seen that 7 (23.3 %) samples in primary endodontic infection group and 24 (80 %) samples in persistent endodontic infection group were positive for D. invisus. On the other hand, 11 (36.6 %) patients of primary endodontic infection showed positivity for O. uli in comparison to 9 (30 %) of persistent endodontic infection. Conclusion: The results from the present study showed efficient amplification of both O. uli and D. invisus in a single-step PCR. Hence, we conclude that the modified protocol used here with taq polymerase enzyme offers a faster and cheaper alternative to nested PCR without compromising the quality of amplification process.

scholarly journals Development of a Specific Polymerase Chain Reaction System for the Detection of Rice Orange Leaf Phytoplasma

Plant Disease ◽  
2020 ◽  
Vol 104 (2) ◽  
pp. 521-526
Author(s):  
Zhiyi Wang ◽  
Yingzhi Zhu ◽  
Zhanbiao Li ◽  
Xin Yang ◽  
Tong Zhang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Nested Pcr ◽  
False Positive ◽  
Southern China ◽  
Reaction System ◽  
Universal Primers ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Orange Leaf ◽  
Polymerase Chain

Rice orange leaf disease (ROLD), caused by rice orange leaf phytoplasma (ROLP), is transmitted by leafhopper vectors Recilia dorsalis and Nephotettix cinticeps. ROLD severely devastates rice production in Asia. Accurate detection of the pathogen is important for disease management. Current nested polymerase chain reaction (nested PCR) method using phytoplasma universal primers is widely used to detect phytoplasmas; however, it has shortcoming of inconvenience and inaccuracy, for it needs two round of PCR reactions and could produce false positive results due to nontarget amplification. In this study, we developed a PCR assay using a set of primers designed based on the ROLP genome sequence to amplify house-keeping gene FtsH-1 in rice and leafhopper vector samples. This method is simple and rapid, and its sensitivity up to 10 pg/μl of total ROLP DNA. It also minimizes the false positive problem produced by nested PCR. This method was used to survey the geographic distribution of ROLD in southern China from 2016 to 2018. The results showed that the distribution areas and vector carrying rate of ROLD had gradually increased.

Application of polymerase chain reaction for detection of goats' milk adulteration by milk of cow

2001 ◽  
Vol 68 (2) ◽  
pp. 333-336 ◽  
Author(s):  
JACEK BANIA ◽  
MACIEJ UGORSKI ◽  
ANTONI POLANOWSKI ◽  
ERYK ADAMCZYK
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Analysis ◽  
Marker Gene ◽  
Meat Products ◽  
Universal Primers ◽  
Animal Origin ◽  
Chain Reaction ◽  
Specific Primers ◽  
Single Stranded Conformational Polymorphism ◽  
Polymerase Chain

Numerous methods based on DNA analysis have been employed in the food industry to monitor adulterations of food products of animal origin. Among them the most frequently used are: polymerase chain reaction (PCR) amplification of a marker gene fragment(s) with universal primers, or amplification of DNA with species-specific primers. PCR-products of different origin can be discriminated by size, restriction fragment length polymorphism (RFLP) or single stranded conformational polymorphism (SSCP) analysis. These methods have been used for identification, and differentiation between, the animal origins of raw or heat-treated meat and meat products (Chikuni et al. 1994; Meyer et al. 1994, 1995; Zehner et al. 1998; Behrens et al. 1999; Guoli et al. 1999; Hopwood et al. 1999; Matsunaga et al. 1999; Wolf et al. 1999). These approaches are also applicable to the analysis of dairy products. However, adulterations of goats' milk and its products are traditionally tested by immunological and/or electrophoretic methods (Amigo et al. 1992; Levieux & Venien, 1994; Mimmo & Pagani, 1998). So far, only a few DNA-based techniques designed to detect the presence of bovine DNA in goats' milk have been described (Plath et al. 1997; Branciari et al. 2000). This paper presents a one-step PCR procedure for detection of adulteration of goats' milk with cows' milk. The method, employing bovine-specific primers for amplification of a 274 bp fragment of cytochrome b DNA, seems to be simple, fast, specific and sensitive.

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