The use of quantitative polymerase chain reaction for the detection and enumeration of the harmful algaAureococcus anophagefferensin environmental samples along the United States East Coast

2003 ◽  
Vol 1 (1) ◽  
pp. 92-102 ◽  
Author(s):  
Linda C. Popels ◽  
S. Craig Cary ◽  
David A. Hutchins ◽  
Rachel Forbes ◽  
Frances Pustizzi ◽  
...  
Keyword(s):  
United States ◽  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
East Coast ◽  
Quantitative Polymerase Chain Reaction ◽  
The United States ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Transmission of the Citrus Variegated Chlorosis Bacterium Xylella fastidiosa with the Sharpshooter Oncometopia nigricans

Plant Disease ◽  
2002 ◽  
Vol 86 (11) ◽  
pp. 1237-1239 ◽  
Author(s):  
R. H. Brlansky ◽  
V. D. Damsteegt ◽  
J. S. Hartung
Keyword(s):  
United States ◽  
Polymerase Chain Reaction ◽  
Xylella Fastidiosa ◽  
The United States ◽  
Reliable Indicator ◽  
Symptom Development ◽  
Chain Reaction ◽  
Citrus Variegated Chlorosis ◽  
Specific Polymerase Chain Reaction ◽  
Polymerase Chain

Citrus variegated chlorosis (CVC) is an economically important, destructive disease in Brazil and is caused by the bacterium Xylella fastidiosa Wells. The bacterium has been found to be transmitted in Brazil by sharpshooter leafhoppers (Cicadellidae). Sharpshooters are present in most citrus growing areas of the United States. The sharpshooter leafhopper, Oncometopia nigricans Walker, frequently is found feeding on citrus in Florida. This sharpshooter transmits the X. fastidiosa strains that cause Pierce's disease of grape and ragweed stunt. Research was initiated to determine if O. nigricans was capable of vectoring the X. fastidiosa that causes CVC. In 59 different transmission tests, using 1 to 57 insects per test, transmission of the bacterium was observed 12 times (20.3%). Symptom development in the greenhouse was not a reliable indicator of transmission. Transmission was verified by specific polymerase chain reaction-based assays. Individual insects were able to transmit the bacterium. This information on sharpshooter transmission of CVC is needed to assess the threat posed by the CVC disease to the citrus industries in the United States.

scholarly journals Two Distinct Genotypes of Spissistilus festinus (Say, 1830) (Hemiptera, Membracidae) in the United States Revealed by Phylogenetic and Morphological Analyses

Insects ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 80
Author(s):  
Elizabeth Cieniewicz ◽  
Victoria Poplaski ◽  
Melina Brunelli ◽  
Jason Dombroskie ◽  
Marc Fuchs
Keyword(s):  
United States ◽  
Polymerase Chain Reaction ◽  
Southeastern United States ◽  
Variable Region ◽  
The United States ◽  
Coi Gene ◽  
Its2 Region ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Spissistilus Festinus

Spissistilus festinus (Say, 1830) (Hemiptera: Membracidae) is a frequent pest of leguminous crops in the Southern United States, and a vector of grapevine red blotch virus. There is currently no information on the genetic diversity of S. festinus. In this study, populations of S. festinus were collected in 2015–2017 from various crops and geographic locations in the United States, and fragments of the mitochondrial cytochrome C oxidase 1 (mt-COI) gene and the nuclear internal transcribed spacer 2 (ITS2) region were characterized by polymerase chain reaction and sequencing. Maximum-likelihood and Bayesian analyses of the mt-COI and ITS2 sequences yielded similar phylogenetic tree topologies, revealing two distinct genetic S. festinus lineages with all of the specimens from California comprising one phylogenetic clade, alongside a single GenBank entry from Arizona, and all specimens from the Southeastern United States comprising a statistically-supported distinct clade, regardless of host and year of collection. The mt-COI gene fragment showed up to 10.8% genetic distance between the two phylogenetic clades. These results suggest the existence of two genotypes within S. festinus in the United States. The only distinct morphological trait between the two genotypes was a less elevated pronotum in the representative specimens from California, compared to the representative specimens from the Southeastern United States. Since this phenotypic feature is inconspicuous, a diagnostic polymerase chain reaction targeting a variable region of the mt-COI fragment was developed to reliably distinguish between the specimens of the two genotypes of S. festinus and to facilitate their specific identification.

Case Report: Molecular Diagnosis of Cystoisospora belli in a Severely Immunocompromised Patient with HIV and Kaposi Sarcoma

2021 ◽  
Author(s):  
Roumen Borilov Iordanov ◽  
Lauren M. Leining ◽  
Meng Wu ◽  
Galant Chan ◽  
Andrew R. DiNardo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Immunocompromised Patient ◽  
Quantitative Polymerase Chain Reaction ◽  
Kaposi Sarcoma ◽  
The United States ◽  
Watery Diarrhea ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Diagnostic Modalities ◽  
Relapsed Disease

Diarrhea in an immunocompromised patient has a broad infectious differential. Diagnosis is difficult despite advances in diagnostic modalities. We report a case of a 45-year-old Nigerian woman who immigrated to the United States 2 years ago. She presented to the hospital with gastrointestinal bleeding, newly diagnosed HIV, and disseminated Kaposi sarcoma. During hospitalization, the patient had an onset of watery diarrhea and high eosinophilia. Subsequent stool analysis using multi-parallel real-time quantitative polymerase chain reaction for 13 parasites was positive for Cystoisospora belli. The patient was treated with trimethoprim–sulfamethoxazole, but had relapsed disease when her antibiotics were stopped prematurely. After restarting trimethoprim–sulfamethoxazole, her diarrhea and eosinophilia improved, and she had undetectable Cystoisospora belli DNA on repeat stool quantitative polymerase chain reaction. This case highlights the importance of a thorough workup for diarrhea, including parasites, especially for immunocompromised patients. Antibiotic prophylaxis is recommended in patients with Cystoisospora belli and HIV/AIDS.

scholarly journals Ralstonia solanacearum Race 3, Biovar 2, the Causal Agent of Brown Rot of Potato, Identified in Geraniums in Pennsylvania, Delaware, and Connecticut

Plant Disease ◽  
2003 ◽  
Vol 87 (4) ◽  
pp. 450-450 ◽  
Author(s):  
S. H. Kim ◽  
T. N. Olson ◽  
N. W. Schaad ◽  
G. W. Moorman
Keyword(s):  
United States ◽  
Polymerase Chain Reaction ◽  
Bacterial Wilt ◽  
Ralstonia Solanacearum ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Carbohydrate Utilization ◽  
Tetrazolium Chloride ◽  
Polymerase Chain

The Plant Disease Diagnostic Laboratory of the Pennsylvania Department of Agriculture received diseased geranium (Pelargonium × hortorum) samples from several Pennsylvania (PA) greenhouses in 1999 and 2000 and from one Delaware (DE) greenhouse in 1999. Originating from Guatemala, plants exhibited yellowing, wilting, stunting, and bacterial oozing from the vascular tissues. Isolations on yeast dextrose-CaCO3 (YDC) and triphenyl-tetrazolium-chloride (TTC) agars resulted in off-white mucoid colonies and white, fluidal colonies with pink centers, respectively. Such colonies are typical of Ralstonia solanacearum (1). The disease was similar to a bacterial wilt of geranium caused by an unidentified biovar of R. solanacearum (3). Preliminary tests using Biolog MicroLog 3 (Hayward, Ca; 4.01A) and enzyme-linked immunosorbent assay (ELISA) (Agdia Inc., Elkhart, IN; BRA 33900/0500) identified the organism as R. solanacearum. For pathogenicity tests, a 10-μl droplet of water suspension containing 1 × 106 CFU of each of five geranium strains (PDA 22056-99, 81849-99, 81862-99, 51032-00, and 64054-00) per milliliter was placed on a stem wound made by cutting off the terminal growth of each of 4 6-leaf stage plants of geranium ‘Orbit Scarlet’, tomato ‘Rutgers’, potato ‘Russet Norkotah’, and eggplant ‘Black Beauty’ in a growth chamber at 28°C, 86% relative humidity, and 12 h light/dark cycle. Water was included as a control. The five strains caused severe yellowing and wilting within 10 days. Colonies typical of R. solanacearum were reisolated from symptomatic tissue on YDC and TTC. To determine the specific biovar, 20 pathogenic geranium strains from PA and DE plus a strain of R. solanacearum originally isolated from a geranium plant of Guatemalan origin received from Connecticut in 1995 were grown up to 28 days on Ayers mineral medium supplemented with a 1% final concentration of D-cellobiose, dextrose, meso-inositol, lactose, maltose, D-ribose, trehalose, mannitol, sorbitol, or dulcitol (1). Acid was produced by 21 test strains from the first five carbohydrates only. Such carbohydrate utilization is typical of bv 2 (1). Bv 2 identification was confirmed by real-time polymerase chain reaction using bv 2-specific primers and probes (N. Schaad, unpublished) designed from a bv 2-specific DNA fragment (2). All tested strains were positive using ELISA. In contrast, strains of bv 2 from geraniums in Wisconsin and South Dakota were reported to be negative using ELISA (4). From our results, it appears that bv 2 was introduced into the United States on geraniums from Guatemala in 1995 and 1999. This cool climate bv 2, a regulated agent by the Agricultural Bioterrorism Protection Act of 2002, has caused extensive crop loss in potatoes in Europe, but has not been found in potatoes in the United States. References: (1) T. P. Denny and A. C. Hayward. Ralstonia solanacearum. Pages 151–174 in: Lab Guide for Identification of Plant Pathogenic Bacteria. N. W. Schaad et al. eds. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001. (2) M. Fagen et al. Development of a diagnostic test based on the polymerase chain reaction (PCR) to identify strains of R. solanacearum exhibiting the Biovar 2 genotype. Pages 34–43 in: Bacterial Wilt Disease: Molecular and Ecological Aspects. P. H. Prior et al. eds. Springer-Verlag, Berlin, 1998. (3) D. L. Strider et al. Plant Dis. 65:52, 1981. (4) L. Williamson et al. (Abstr.) Phytopathology 91 (Suppl.):S95, 2001.

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