Digenetic trematodes,Acanthatriumsp. andLecithodendriumsp., as vectors ofNeorickettsia risticii, the agent of Potomac horse fever

2003 ◽  
Vol 77 (4) ◽  
pp. 335-339 ◽  
Author(s):  
N. Pusterla ◽  
E.M. Johnson ◽  
J.S. Chae ◽  
J.E. Madigan
Keyword(s):  
Aquatic Insects ◽  
Preliminary Evidence ◽  
Tachycineta Bicolor ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Nested Polymerase Chain Reaction ◽  
Digenetic Trematodes ◽  
Potomac Horse Fever ◽  
The Family ◽  
Polymerase Chain

AbstractNeorickettsia(formerlyEhrlichia)risticii, the agent of Potomac horse fever (PHF), has been recently detected in trematode stages found in the secretions of freshwater snails and in aquatic insects. Insectivores, such as bats and birds, may serve as the definitive host of the trematode vector. To determine the definitive helminth vector, five bats (Myotis yumanensis) and three swallows (Hirundo rustica,Tachycineta bicolor) were collected from a PHF endemic location in northern California. Bats and swallows were dissected and their major organs examined for trematodes and forN. risticiiDNA using a nested polymerase chain reaction (PCR) assay. Adult digenetic trematodes,Acanthatriumsp. and/orLecithodendriumsp., were recovered from the gastrointestinal tract of all bats and from one swallow. The intestine of three bats, the spleen of two bats and one swallow as well as the liver of one swallow tested PCR positive forN. risticii. From a total of seven pools of identical digenetic trematodes collected from single hosts, two pools ofAcanthatriumsp. and one pool ofLecithodendriumsp. tested PCR positive. The results of this investigation provide preliminary evidence that at least two trematodes in the family Lecithodendriidae are vectors ofN. risticii. The data also suggest that bats and swallows not only act as a host for trematodes but also as a possible natural reservoir forN. risticii.

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 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 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.

scholarly journals Identification of Leptospira in Patients With Renal Failure Using Serological, Molecular and Pathological Based Techniques, in Shiraz, Iran

2022 ◽  
Author(s):  
Mostafa Sayyadi ◽  
Saeid Hosseinzadeh ◽  
Gholamreza Abdollahpour ◽  
Seyed Shahram Shekarforoush ◽  
Azadeh Samiei ◽  
...  
Keyword(s):  
Staining Method ◽  
Pathogenic Species ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Serum Samples ◽  
Biochemical Tests ◽  
Nested Polymerase Chain Reaction ◽  
Renal Disorders ◽  
Silver Staining Method ◽  
Polymerase Chain

Abstract Leptospirosis is a relatively rare bacterial infection that affects people and animals caused by pathogenic species of Leptospira. The present study was conducted using nested polymerase chain reaction (PCR), microscopic agglutination test (MAT) and hematological and biochemical tests on 200 blood samples of renal disorders patients in Shiraz, Iran. Also nested-PCR assay and Warthin-Starry (WS) silver staining method was performed on 30 nephrectomised kidney sample. The frequency of pathogenic species of Leptospira infection in patients with renal disorders was 20 % and this infection was significantly correlated with BUN, anemia, RDW, MCV, MCH and hemoglobin levels (P < 0.01). MAT analysis showed that serum samples had positive titers against L. Grippotyphosa (13 samples), L. Ballum (6 sample), L. Pomona (3 samples), L. Canicola (2 samples), L. Icterohaemorrhagiae (1 sample) and L. Hardjo (1 sample) serovars. Twenty-three percent of the kidney samples from the patients with pyelonephritis were infected with the pathogenic species of Leptospira. This study showed that pathogenic Leptospira serovars are present in this area and in patients with renal disorders more attention should be paid to this zoonotic disease.

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

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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