scholarly journals A Novel Nested Polymerase Chain Reaction (n-PCR) Assay for Identifying Sorghum nitidum

2011 ◽  
Vol 3 (4) ◽  
pp. 143-146
Author(s):  
Shasha WEI ◽  
Zhirui DENG ◽  
Liping YIN ◽  
Jianping YI ◽  
Renqi WU ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleotide Sequence ◽  
Sorghum Bicolor ◽  
Plant Protection ◽  
Sorghum Halepense ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Nested Polymerase Chain Reaction ◽  
Field Inspection ◽  
Polymerase Chain

This work developed a novel nested polymerase chain reaction (n-PCR) assay to identify Sorghum nitidum (S. nitidum). It has been designed a set of specific n-PCR inner primers Snit5/Snit2 and outer primers Nout1/Nout2 based on a conserved nucleotide sequence of adh1-like gene of S. nitidum. Fourteen samples of sorghum were used to investigate the specificity of the primers and the n-PCR assay. The result showed that 9 samples of S. nitidum displayed a positive strong, specific amplified band at ~873 bp in gel spectra, while other relatives, including Sorghum halepense, Sorghum almum, Sorghum bicolor, Sorghum propinum and Sorghum sudanse exhibited negative amplifications. This assay was able to specifically identify S. nitidum fast and effectively, which could be applied widely in field inspection, agriculture production and plant protection.

scholarly journals Sensitivity of the nested-polymerase chain reaction (PCR) assay for Brugia malayi and significance of ‘free’ DNA in PCR-based assays

2000 ◽  
Vol 30 (11) ◽  
pp. 1177-1179 ◽  
Author(s):  
Janet Cox-Singh ◽  
Andrea S Pomrehn ◽  
Nathan D Wolfe ◽  
Hasan A Rahman ◽  
Huong-Ying Lu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Brugia Malayi ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Nested Polymerase Chain Reaction ◽  
Free Dna ◽  
Polymerase Chain

scholarly journals Evaluation and Use of a Nested Polymerase Chain Reaction Assay in Cats Experimentally Infected with Bartonella Henselae Genotype I and Bartonella Henselae Genotype II

2001 ◽  
Vol 13 (4) ◽  
pp. 312-322 ◽  
Author(s):  
Alma F. Roy ◽  
Richard E. Corstvet ◽  
Ronald A. Tapp ◽  
Kathy L. O'Reilly ◽  
Hollis U. Cox
Keyword(s):  
Polymerase Chain Reaction ◽  
Bartonella Henselae ◽  
Turnaround Time ◽  
Nucleic Acid Amplification ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Nested Polymerase Chain Reaction ◽  
Genotype I ◽  
Polymerase Chain ◽  
Genotype Ii

Cats have been shown to be infected with Bartonella henselae genotype I, B. henselae genotype II, and B. clarridgeiae. Feline bartonellosis infections and the strains involved in these infections are important in both veterinary and human medicine. Nucleic acid amplification methods such as polymerase chain reaction (PCR) are being used in both research and diagnostics as tools for understanding many infectious diseases. Bartonella bacteremia in cats is detected by blood culture; however, because of the limitations of culture (delayed turnaround time and sensitivity limits), PCR may be a more efficient method for identifying infected cats. Three distinct PCR assays that could differentiate among B. henselae genotype I, B. henselae genotype II, and B. clarridgeiae were developed and used to detect as few as 3.2 organisms. Fourteen cats experimentally infected with B. henselae genotype I and B. henselae genotype II were followed by bacterial culture and PCR through the course of infection, including periods of primary and relapsing bacteremia. The PCR assay was positive in 11 of the 14 cats for periods of 1–9 weeks after culture became negative. Of the 223 blood specimens that were culture negative, the PCR assay was positive in 38 (17%) of the specimens. Two of the 14 cats developed relapsing bacteremia. The 2 B. henselae genotypes were amplified in the cats and the bacteremic phase of these infections as determined by PCR lasted for a longer period than previously determined by culture. Using laboratory assays such as PCR to understand the strains involved in feline bartonellosis and the course of the infection is important in the understanding of these zoonotic agents.

scholarly journals Interception of ‘Candidatus Phytoplasma solani’ in Brazil in apple tree propagative material imported from France

2018 ◽  
Vol 84 (0) ◽  
Author(s):  
Barbara Eckstein ◽  
Stephanie Regina Alves Botelho ◽  
Mayara Luana Coelho Alves de Oliveira ◽  
Fernanda Rausch Fernandes ◽  
Márcio Martinello Sanches
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleotide Sequence ◽  
Apple Tree ◽  
Plant Protection ◽  
Apple Rootstock ◽  
Chain Reaction ◽  
Phytoplasma Infection ◽  
Polymerase Chain ◽  
Exotic Pest ◽  
Grafting Onto

ABSTRACT: Apple plants from France introduced in Brazil for research purposes were subjected to a phytosanitary analysis at the Plant Quarantine Laboratory of Embrapa Genetic Resources and Biotechnology (Cenargen). After grafting onto healthy apple rootstock, some plants showed phytoplasma-infection symptoms. Polymerase Chain Reaction (PCR) tests yielded DNA fragments of the expected size for phytoplasmas. DNA sequencing revealed an identity of the 16S rDNA nucleotide sequence of 98-99% with ‘Candidatus Phytoplasma solani’. This phytoplasma species is responsible for losses in European apple orchards and has not been reported in Brazil. According to the Federal Legislation on Plant Protection, the plants were incinerated to avoid the introduction of this exotic pest in Brazil.

scholarly journals Development of a Nested Polymerase Chain Reaction Assay for Detection of Colletotrichum acutatum on Symptomless Strawberry Leaves

Plant Disease ◽  
2008 ◽  
Vol 92 (12) ◽  
pp. 1655-1661 ◽  
Author(s):  
O. Pérez-Hernández ◽  
M. H. Nam ◽  
M. L. Gleason ◽  
H. G. Kim
Keyword(s):  
Polymerase Chain Reaction ◽  
Colletotrichum Acutatum ◽  
Tween 20 ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Nested Polymerase Chain Reaction ◽  
Plastic Bags ◽  
Reliable Diagnosis ◽  
Polymerase Chain ◽  
Composite Samples

A nested polymerase chain reaction (PCR) assay was developed for detection of Colletotrichum acutatum on symptomless strawberry leaves. In pure culture, the assay detected as little as 1.0 fg of DNA extracted from mycelium and as few as 1.5 conidia ml–1 when conidial suspensions were sonicated. On detached inoculated leaves, three alternative protocols to dislodge the pathogen were assessed: (i) immersion of whole leaves in 0.05% Tween 20 and manual agitation in plastic bags for 1 min (A); (ii) immersion in Tween 20, sonication for 30 min, then agitation for 1 min (SA); and (iii) freezing for 3 h, incubation for 2 days at 27°C, immersion in Tween 20, then sonication for 30 min and agitation for 1 min (FISA). Each method removed significantly (P ≤ 0.05) more conidia from leaves than the nontreated control; however, removal of appressoria did not vary among assays. In composite samples of noninoculated and inoculated (1.5 ×103 conidia ml–1) strawberry leaves, the nested PCR assay using the FISA protocol detected C. acutatum in as few as 1 infested leaf in 50 noninfested leaves. In a strawberry field, the assay detected the presence of C. acutatum in samples of asymptomatic strawberry leaves, showing potential as a powerful tool for reliable diagnosis of the pathogen in the field.

Detection of hepatitis C viral sequences in serum by ‘nested’ polymerase chain reaction (PCR) and a commercial single-round PCR assay

1995 ◽  
Vol 4 (3) ◽  
pp. 239-250 ◽  
Author(s):  
Harald H. Kessler ◽  
Brigitte Santner ◽  
Florian Umlauft ◽  
Martina Urbanek ◽  
Max Kronawetter ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Hepatitis C ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Viral Sequences

scholarly journals Stolbur Phytoplasma Transmission to Maize by Reptalus panzeri and the Disease Cycle of Maize Redness in Serbia

2009 ◽  
Vol 99 (9) ◽  
pp. 1053-1061 ◽  
Author(s):  
J. Jović ◽  
T. Cvrković ◽  
M. Mitrović ◽  
S. Krnjajić ◽  
A. Petrović ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sorghum Halepense ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Disease Cycle ◽  
Maize Roots ◽  
Infected Adult ◽  
Polymerase Chain ◽  
Border Areas ◽  
Stolbur Phytoplasma

Maize redness (MR), induced by stolbur phytoplasma (‘Candidatus Phytoplasma solani’, subgroup 16SrXII-A), is characterized by midrib, leaf, and stalk reddening and abnormal ear development. MR has been reported from Serbia, Romania, and Bulgaria for 50 years, and recent epiphytotics reduced yields by 40 to 90% in South Banat District, Serbia. Potential vectors including leafhoppers and planthoppers in the order Hemiptera, suborder Auchenorrhyncha, were surveyed in MR-affected and low-MR-incidence fields, and 33 different species were identified. Only Reptalus panzeri populations displayed characteristics of a major MR vector. More R. panzeri individuals were present in MR-affected versus low-MR fields, higher populations were observed in maize plots than in field border areas, and peak population levels preceded the appearance of MR in late July. Stolbur phytoplasma was detected in 17% of R. panzeri adults using nested polymerase chain reaction but not in any other insects tested. Higher populations of R. panzeri nymphs were found on maize, Johnsongrass (Sorghum halepense), and wheat (Triticum aestivum) roots. Stolbur phytoplasma was detected in roots of these three plant species, as well as in R. panzeri L3 and L5 nymphs. When stolbur phytoplasma-infected R. panzeri L3 nymphs were introduced into insect-free mesh cages containing healthy maize and wheat plants, 89 and 7%, respectively, became infected. These results suggest that the MR disease cycle in South Banat involves mid-July transmission of stolbur phytoplasma to maize by infected adult R. panzeri. The adult R. panzeri lay eggs on infected maize roots, and nymphs living on these roots acquire the phytoplasma from infected maize. The nymphs overwinter on the roots of wheat planted into maize fields in the autumn, allowing emergence of phytoplasma-infected vectors the following July.

Clonorchis sinensis: Development and evaluation of a nested polymerase chain reaction (PCR) assay

2007 ◽  
Vol 115 (3) ◽  
pp. 291-295 ◽  
Author(s):  
Ammini Parvathi ◽  
H. Sanath Kumar ◽  
B. Kenchanna Prakasha ◽  
Jieyuan Lu ◽  
Xuenian Xu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Clonorchis Sinensis ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain

Nested polymerase chain reaction for detection of pathogenic leptospires

2006 ◽  
Vol 52 (8) ◽  
pp. 747-752 ◽  
Author(s):  
Sandra Denize Dorneles Jouglard ◽  
Simone Simionatto ◽  
Fabiana Kommling Seixas ◽  
Fernanda Lima Nassi ◽  
Odir Antônio Dellagostin
Keyword(s):  
Polymerase Chain Reaction ◽  
Nested Pcr ◽  
Early Stage ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Nested Polymerase Chain Reaction ◽  
Pathogenic Leptospira ◽  
Polymerase Chain ◽  
Low Sensitivity

Leptospirosis is a widespread zoonosis caused by pathogenic members of the genus Leptospira that has a great impact on human and veterinary public health. Early diagnosis of leptospirosis is important because severe lepto spiral infection can have a fulminant course. The available serological techniques for the diagnosis of leptospirosis have low sensitivity during the early stage of the disease. Efforts are being made to develop simpler, effective, efficient, and inexpensive diagnostic methods. In this work, we first evaluate a polymerase chain reaction (PCR) based method for diagnosis of leptospirosis. Primers were designed to amplify a 264 bp region within the lipL32 gene that is conserved among pathogenic Leptospira and absent in nonpathogenic species. The sensitivity and specificity of the assay were evaluated using 7 saprophytic serovars, 37 pathogenic serovars, and 15 other microorganisms. The method was very specific for pathogenic serovars, however, it lacked sensitivity. To enhance the sensitivity, another primer pair was designed to amplify a 183 bp region within the 264 bp region of the lipL32 gene and was used in a nested PCR assay. This approach was much more sensitive than conventional PCR.Key words: leptospirosis, diagnosis, nested PCR, lipL32.

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