scholarly journals Multi-locus sequence typing for species/serovar identification of clinical isolates of Leptospira spp

2021 ◽  
Vol 52 (3) ◽  
Author(s):  
K. Justin Davis ◽  
K. Justin Davis ◽  
K. Justin Davis ◽  
K. Justin Davis ◽  
K. Justin Davis
Keyword(s):  
Solid Medium ◽  
Control Strategies ◽  
Multi Locus Sequence Typing ◽  
Close Monitoring ◽  
Mlst Database ◽  
Chain Reaction ◽  
Allelic Profile ◽  
Pathogenic Leptospira ◽  
Polymerase Chain ◽  
Semi Solid

Leptospirosis is an emerging zoonotic disease endemic in Kerala and close monitoring of emerging serovars is essential to adopt appropriate control strategies. Multi-Locus Sequence Typing (MLST) was reported to be less expensive compared to other cumbersome methods like whole genome sequencing. The present study was conducted to obtain isolates of Leptospira from infected animals and rats and for the identification of serovars using MLST. A total of 205 blood samples (dog, cat, cattle, goat), 43 urine samples (dog, cattle) and post-mortem kidney samples from various animals such as dog (n=12), cattle (n=2) and rat (n=25) were collected and subjected to polymerase chain reaction (PCR) using G1/G2 primers to identify the pathogenic Leptospira. Fifteen samples were found to be positive, these samples when inoculated in the Ellinghausen- McCullough-Johnson-Harris (EMJH) semi-solid medium to obtain ten isolates. These ten isolates were further subjected to secY, icdA and GyraseB PCR and sequenced. The obtained sequences were analysed using BLAST and were fed into specified MLST database of Leptospira scheme-3, the allelic profile and sequence type were generated. The MLST results obtained in the study indicated that the isolates S24 and S33 belonged to serovar Canicola, S40 and 47 were Sejroe and S19, S27, S55, S69 and S71 were Bataviae, Autumnalis, Pomona, Icterohaemorraghiae and Australis, respectively. It was concluded that MLST is a convenient method for identifying leptospiral serovars.

scholarly journals Use of the polymerase chain reaction for detection of Fusarium graminearum in bulgur wheat

2012 ◽  
Vol 32 (1) ◽  
pp. 201-208 ◽  
Author(s):  
Carla Bertechini Faria ◽  
Giovana Caputo Almeida-Ferreira ◽  
Karina Bertechine Gagliardi ◽  
Tatiane Cristina Albuquerque Alves ◽  
Dauri José Tessmann ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Fusarium Graminearum ◽  
Genomic Dna ◽  
Solid Medium ◽  
Food Contamination ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Culture Characteristics ◽  
Mycotoxigenic Fungi ◽  
Polymerase Chain

The detection of mycotoxigenic fungi in foodstuff is important because their presence may indicate the possible associated mycotoxin contamination. Fusarium graminearum is a wheat pathogen and a producer of micotoxins. The polymerase chain reaction (PCR) has been employed for the specific identification of F. graminearum. However, this methodology has not been commonly used for detection of F. graminearum in food. Thus, the objective of the present study was to develop a molecular methodology to detect F. graminearum in commercial samples of bulgur wheat. Two methods were tested. In the first method, a sample of this cereal was contaminated with F. graminearum mycelia. The genomic DNA was extracted from this mixture and used in a F. graminearum specific PCR reaction. The F. graminearum species was detected only in samples that were heavily contaminated. In the second method, samples of bulgur wheat were inoculated on a solid medium, and isolates having F. graminearum culture characteristics were obtained. The DNA extracted from these isolates was tested in F. graminearum specific PCR reactions. An isolate obtained had its trichothecene genotype identified by PCR. The established methodology could be used in surveys of food contamination with F. graminearum.

Leptospirosis: a diagnostic conundrum

2018 ◽  
Vol 48 (4) ◽  
pp. 306-309
Author(s):  
Keerthivasan Gurumurthy ◽  
Harish Belgode Narasimha ◽  
Mukta Wyawahare ◽  
Niranjan Biswal
Keyword(s):  
The Body ◽  
Health Concern ◽  
Andaman Islands ◽  
Public Health Concern ◽  
Chain Reaction ◽  
Pathogenic Leptospira ◽  
Indian States ◽  
Polymerase Chain Reaction Testing ◽  
Laboratory Confirmation ◽  
Polymerase Chain

Leptospirosis is a serious public health concern worldwide. It is highly endemic in the Andaman Islands and its prevalence is increasing in other Indian states. Clinical features are non-specific and diagnosis relies on laboratory confirmation. The gold standard is microscopic agglutination testing, but this is not widely available. Real-time polymerase chain reaction testing of LipL32 antigen provides the earliest detection of pathogenic Leptospira in the body. We found it to be 100% specific, but it should be used in the first 10 days of illness for reliable results.

Genetic transformation of Stella De Oro daylily by particle bombardment

2003 ◽  
Vol 83 (4) ◽  
pp. 873-876 ◽  
Author(s):  
A. N. Aziz ◽  
R. J. Sauvé ◽  
S. Zhou
Keyword(s):  
Southern Blotting ◽  
Basal Medium ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Semi Solid ◽  
Independent Transformation

Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis

scholarly journals Molecular Typing of Human Brucella melitensis Isolated from Patients in Erbil, Iraq

2019 ◽  
Vol 7 (1) ◽  
pp. 1 ◽  
Author(s):  
Bushra K. Amin ◽  
Khulod I. Hassan
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Typing ◽  
Brucella Melitensis ◽  
Control Strategies ◽  
Human Brucellosis ◽  
Kurdistan Region ◽  
Specific Primer ◽  
Molecular Method ◽  
Chain Reaction ◽  
Polymerase Chain

Brucellosis is a reemerging infectious zoonotic disease of worldwide importance. In the Kurdistan Region of Iraq, it is a widely spread disease and remains a challenging health problem. This disease is mainly caused by Brucella melitensis, in human. For confirmation of these isolates, a study was performed, by isolation and molecular typing of Brucella Spp. from human patients in Rizgari Hospital at Erbil city (Iraq), between March 2014 and November 2016. One hundred sixty seven samples of blood collected from patients suspected for brucellosis, one hundred twenty one samples from these were recorded as genus of Brucella, using biochemical test and confirmed by applying polymerase chain reaction (PCR), using genus specific primer for omp31 gene which was specific for B. melitensis. These results support using molecular method that based on PCR as diagnostic test for the control of brucellosis in Erbil. Further studies are needed from different geographical areas of the country with different level of endemicity to plan and execute control strategies against human brucellosis.

scholarly journals Development and validation of real time polymerase chain reaction for detection DNA of pathogenic leptospira

2018 ◽  
Vol 32 (2) ◽  
pp. 565-576
Author(s):  
V. V. Ukhovskyi ◽  
L. M. Muzykina ◽  
I. V. Galka ◽  
V. G. Spyrydonov ◽  
I. I. Antonik ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Chain Reaction ◽  
Pathogenic Leptospira ◽  
Polymerase Chain ◽  
Development And Validation

scholarly journals Identification of Pathogenic Leptospira spp. Carrier Goats Using Polymerase Chain Reaction (PCR)

2017 ◽  
Vol 6 (12) ◽  
pp. 2174-2183 ◽  
Author(s):  
Priti D. Vihol ◽  
Jatin H. Patel ◽  
Jignesh M. Patel ◽  
Vijendra S. Dabas ◽  
Irshadullakhan H. Kalyani ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Pathogenic Leptospira ◽  
Leptospira Spp ◽  
Polymerase Chain

scholarly journals Leptospira Detection in Cats in Spain by Serology and Molecular Techniques

2020 ◽  
Vol 17 (5) ◽  
pp. 1600 ◽  
Author(s):  
Andrea Murillo ◽  
Rafaela Cuenca ◽  
Emmanuel Serrano ◽  
Goris Marga ◽  
Ahmed Ahmed ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Infection ◽  
Molecular Techniques ◽  
Antibody Prevalence ◽  
Chain Reaction ◽  
Pathogenic Leptospira ◽  
Blood Samples ◽  
Polymerase Chain ◽  
Positive Results ◽  
Leptospira Species

Leptospirosis is the most neglected widespread zoonosis worldwide. In Spain, leptospirosis reports in people and animals have increased lately. Cats can become infected with Leptospira, as well as be chronic carriers. The aim of this study was to determine serological antibody prevalence against Leptospira sp., blood DNA, and shedding of DNA from pathogenic Leptospira species in the urine of cats in Spain. Microagglutination tests (MAT) and blood and urine TaqMan real-time polymerase chain reaction (PCR) were performed. Leptospira antibodies were detected in 10/244 cats; with 4.1% positive results (95% confidence interval (CI): 2.1–7.18%). Titers ranged from 1:20 to 1:320 (serovars Ballum; Bataviae; Bratislava; Cynopteri; Grippotyphosa Mandemakers; Grippotyphosa Moskva; Pomona; and Proechimys). The most common serovar was Cynopteri. Blood samples from 1/89 cats amplified for Leptospira DNA (1.12%; 95% CI: 0.05–5.41%). Urine samples from 4/232 cats amplified for Leptospira DNA (1.72%; 95% CI: 0.55–4.10%). In conclusion free-roaming cats in Spain can shed pathogenic Leptospira DNA in their urine and may be a source of human infection. Serovars not previously described in cats in Spain were detected; suggesting the presence of at least 4 different species of pathogenic leptospires in the country (L. borgpetersenii; L. interrogans; L. kirschneri; and L. noguchii).

scholarly journals Temporal and Microspatial Heterogeneity in Transmission Dynamics of Coendemic Plasmodium vivax and Plasmodium falciparum in Two Rural Cohort Populations in the Peruvian Amazon

2020 ◽  
Author(s):  
Angel Rosas-Aguirre ◽  
Mitchel Guzman-Guzman ◽  
Raul Chuquiyauri ◽  
Marta Moreno ◽  
Paulo Manrique ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Plasmodium Vivax ◽  
Control Strategies ◽  
Population Based ◽  
Regional Level ◽  
Control Measures ◽  
Peruvian Amazon ◽  
Chain Reaction ◽  
Seasonal Transmission ◽  
Polymerase Chain

Abstract Background Malaria is highly heterogeneous: its changing malaria microepidemiology needs to be addressed to support malaria elimination efforts at the regional level. Methods A 3-year, population-based cohort study in 2 settings in the Peruvian Amazon (Lupuna, Cahuide) followed participants by passive and active case detection from January 2013 to December 2015. Incidence and prevalence rates were estimated using microscopy and polymerase chain reaction (PCR). Results Lupuna registered 1828 infections (1708 Plasmodium vivax, 120 Plasmodium falciparum; incidence was 80.7 infections/100 person-years (95% confidence interval [CI] , 77.1–84.5). Cahuide detected 1046 infections (1024 P vivax, 20 P falciparum, 2 mixed); incidence was 40.2 infections/100 person-years (95% CI, 37.9–42.7). Recurrent P vivax infections predominated onwards from 2013. According to PCR data, submicroscopic predominated over microscopic infections, especially in periods of low transmission. The integration of parasitological, entomological, and environmental observations evidenced an intense and seasonal transmission resilient to standard control measures in Lupuna and a persistent residual transmission after severe outbreaks were intensively handled in Cahuide. Conclusions In 2 exemplars of complex local malaria transmission, standard control strategies failed to eliminate submicroscopic and hypnozoite reservoirs, enabling persistent transmission.

scholarly journals Allelic profile of Serbian Xanthomonas campestris pv. campestris isolates from cabbage

2020 ◽  
Vol 35 (1) ◽  
pp. 19-26
Author(s):  
Tatjana Popovic ◽  
Aleksandra Jelusic ◽  
Petar Mitrovic ◽  
Renata Ilicic ◽  
Sanja Markovic
Keyword(s):  
Xanthomonas Campestris ◽  
Minimum Spanning Tree ◽  
Housekeeping Genes ◽  
Sequence Type ◽  
Chain Reaction ◽  
Allelic Profile ◽  
Polymerase Chain ◽  
Rot Disease ◽  
Xanthomonas Campestris Pv Campestris

Xanthomonas campestris pv. campestris (Xcc), the causal agent of black rot disease of cabbage (Brassica oleracea var. capitata L.), is one of the most important bacteria which affect proper cabbage growth, leading to head weight and quality losses and thereby drastically reducing its marketing value. The pathogen is genetically diverse, which is evident from the presence of eleven races worldwide and more than thirty combinations of allelic profiles. Therefore, this study aimed to determine the allelic profiles of Serbian cabbage Xcc strains obtained in 2014. The analysis was done on three selected Xcc strains whose DNA was first amplified using polymerase chain reaction (PCR) with four housekeeping genes - P-XdnaK, fyuA, gyrB, and rpoD, then sequenced, and the obtained sequences were finally used to determine allelic profiles. Allelic profiles were determined by comparison with 33 Xcc strains obtained from different hosts and regions, whose allelic profiles had been determined previously. A non-redundant database (NRDB) from the pubMLST was used for allelic profile determination and Phyloviz software for constructing a minimum spanning tree. The obtained allelic profile of all Serbian Xcc cabbage strains was 1, 3, 1, 1 for the P-X-dnaK, fyuA, gyrB and rpoD genes, respectively. This profile is assigned as sequence type 2 (ST2) and it coincides with a Portuguese B. oleracea Xcc strain, CPBF 213, originating from B. oleracea var. costata. No connection between sequence type (ST) and the host was detected.

Sign in / Sign up

Export Citation Format

Share Document