scholarly journals Influence of Selective Agents (EMJH-STAFF), Sample Filtration and pH on Leptospira Interrogans Serovar Icterohaemorrhagiae Cultivation and Isolation from Swine Urine

2021 ◽  
Vol 8 (6) ◽  
pp. 90
Author(s):  
Romana Steinparzer ◽  
Tamara Mair ◽  
Christine Unterweger ◽  
Adi Steinrigl ◽  
Friedrich Schmoll
Keyword(s):  
Diagnostic Tests ◽  
Urine Samples ◽  
Epidemiological Surveillance ◽  
Leptospira Interrogans ◽  
Sample Storage ◽  
Clinical Specimens ◽  
Negative Results ◽  
Swine Urine ◽  
Selective Agents ◽  
Phosphate Buffered Saline

Leptospira spp. cause the zoonotic disease leptospirosis, which occurs in numerous mammalians worldwide. Isolation is still important for serotyping and genotyping of Leptospira, which in turn is essential for epidemiological surveillance of leptospirosis and the development of diagnostic tests and vaccines. However, isolation of Leptospira from clinical specimens is inherently insensitive. This study was conducted to examine the influence of selective agents, sample filtration, sample pH and the use of phosphate buffered saline (PBS) buffer for sample storage to improve the success of cultivation and isolation of Leptospira interrogans serovar Icterohaemorrhagiae from swine urine. EMJH (Ellinghausen McCullough, Johnson and Harris) medium including the selective agents sulfamethoxazole, trimethoprim, amphotericin, fosfomycin and 5-fluorouracil (STAFF) increased the success of Leptospira isolation from spiked swine urine samples. Sample filtration yielded only negative results. Isolation in EMJH-STAFF was successful from swine urine with a density as low as 104 Leptospira/mL, and urine with pH ≤ 7 impaired the cultivation rate. Cultivation and isolation were not improved by the addition of PBS to spiked urine samples prior to storage for 24 h at 4 °C. The results of the study demonstrate that cultivation and isolation of leptospires from swine urine can be improved by enhanced methods.

Chronic Wasting Disease Options of Surveillance in Carpathian Cervids: from the Identification of Characteristic Microscopic Pathologic Changes to Prionic Seeded Conversion and Amplification Assays

2001 ◽  
Vol 71 (3) ◽  
pp. 480-486
Author(s):  
Florica Barbuceanu ◽  
Stelian Baraitareanu ◽  
Stefania-Felicia Barbuceanu ◽  
Gabriel Predoi
Keyword(s):  
Enzyme Immunoassay ◽  
Diagnostic Tests ◽  
Chronic Wasting Disease ◽  
Rapid Diagnostic Tests ◽  
Diagnostic Methods ◽  
Wasting Disease ◽  
Western Immunoblotting ◽  
Negative Results ◽  
Antigen Capture ◽  
Confirmatory Tests

This paper describes the current diagnostic methods of Chronic Wasting Disease (CWD) in cervides used between 2013 and 2017 in Romania. The active surveillance of CWD involves the targeted groups screening by using rapid diagnostic tests (e.g., antigen capture enzyme immunoassay). If the first test does not provide certain negative results, then the confirmatory methods have been used, i.e. histopathology, immunohistochemistry and Western immunoblotting. These tests did not lead to the detection of CWD prions (PrPCWD) in Romania. This may be due to the absence or insufficient quantity of PrPCWD in samples, below the threshold of confirmatory tests.

scholarly journals Detection of three pandemic causing coronaviruses from non-respiratory samples: systematic review and meta-analysis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chandan Mishra ◽  
Suneeta Meena ◽  
Jitendra Kumar Meena ◽  
Suman Tiwari ◽  
Purva Mathur
Keyword(s):  
Systematic Review ◽  
Detection Rate ◽  
Meta Analysis ◽  
Total Sample ◽  
Urine Samples ◽  
Pathogenic Potential ◽  
Clinical Specimens ◽  
Qrt Pcr ◽  
Corona Virus ◽  
Positive Rate

AbstractSARS-CoV-2 has posed an unprecedented challenge to the world. Pandemics have been caused previously by viruses of this family like Middle East Respiratory Corona Virus (MERS CoV), Severe Acute Respiratory Syndrome Corona Virus (SARS CoV). Although these viruses are primarily respiratory viruses, but they have been isolated from non-respiratory samples as well. Presently, the detection rate of SARS‐CoV‐2 RNA from different clinical specimens using Real Time Reverse Transcriptase Polymerized Chain Reaction (qRT‐PCR) after onset of symptoms is not yet well established. Therefore, the aim of this systematic review was to establish the profile of detecting SARS‐CoV‐2, MERS CoV, SARS CoV from different types of clinical specimens other than the respiratory using a standard diagnostic test (qRT‐PCR). A total of 3429 non-respiratory specimens were recorded: SARS CoV (total sample—802), MERS CoV (total sample—155), SARS CoV-2 (total sample—2347). Out of all the samples studied high positive rate was seen for saliva with 96.7% (14/14; 95% CI 87.6–100.0%) for SARS CoV and 57.5% (58/250; 95% CI − 1.2 to 116.2%) for SARS CoV-2, while low detection rate in urine samples for SARS CoV-2 with 2.2% (8/318; 95% CI 0.6–3.7%) and 9.6% (12/61; 95% CI − 0.9 to 20.1%) for SARS CoV but there was relatively higher positivity in urine samples for MERS CoV with detection rate of 32.4% (2/38; 95% CI − 37.3 to 102.1%). In Stool sample positivity was 54.9% (396/779; 95% CI 41.0–68.8%), 45.2% (180/430; 95% CI 28.1–62.3%) and 34.7% (4/38; 95% CI − 29.5 to 98.9%) for SARS CoV-2, MERS CoV, and SARS CoV, respectively. In blood sample the positivity was 33.3% (7/21; 95% CI 13.2–53.5%), 23.7% (42/277; 95% CI 10.5–36.9%) and 2.5% (2/81; 95% CI 0.00–5.8%) for MERS CoV, SARS CoV-2 and SARS CoV respectively. SARS‐CoV‐2 along with previous two pandemic causing viruses from this family, were highly detected stool and saliva. A low positive rate was recorded in blood samples. Viruses were also detected in fluids along with unusual samples like semen and vaginal secretions thus highlighting unique pathogenic potential of SARS‐CoV‐2.

scholarly journals The Positive Predictive Value of Diagnostic Ultrasound for Occult Herniae

2006 ◽  
Vol 88 (2) ◽  
pp. 165-167 ◽  
Author(s):  
M Bradley ◽  
J Morgan ◽  
B Pentlow ◽  
A Roe
Keyword(s):  
Positive Predictive Value ◽  
Predictive Value ◽  
Diagnostic Tests ◽  
Diagnostic Ultrasound ◽  
Pain Clinic ◽  
Diagnostic Problem ◽  
Clinical Criteria ◽  
Negative Results ◽  
Positive Results

INTRODUCTION The aim of this study is to ascertain the accuracy of diagnostic ultrasound in the assessment of the occult abdominal and groin herniae. The authors have previously demonstrated its efficacy in diagnosing the type of clinical groin herniae but occult herniae provide a further diagnostic problem. PATIENTS AND METHODS A total of 113 consecutive patients were referred prospectively for ultrasound examinations with clinically suspected occult herniae. All positive scans were offered surgery whilst the negative results were offered further imaging or other diagnostic tests depending on the clinical criteria. The end point for negative scans was based on 18-month follow-up or resolution of symptoms. RESULTS Overall, 59 scans showed positive results for herniae and 56 of these had surgery. In the other three patients, two refused an operation, and one had no hernia detected at operation. In the remaining 57 scans, ultrasound offered alternative soft tissue diagnoses in 23 patients and surgical/endoscopic diagnoses accounted for a further 8 patients. CONCLUSIONS Ultrasound offered a diagnosis for the symptomology in 82 patients (70.6%) of which 59 were herniae. The positive predictive value for hernia is 98.3%. Twenty-six patients with no diagnosis or confirmation of herniae on follow-up showed symptom resolution in 22 cases, and four patients were treated by the pain clinic.

Determination of cannabinoids in oral fluid and urine of “light cannabis” consumers: a pilot study

2018 ◽  
Vol 57 (2) ◽  
pp. 238-243 ◽  
Author(s):  
Roberta Pacifici ◽  
Simona Pichini ◽  
Manuela Pellegrini ◽  
Roberta Tittarelli ◽  
Flaminia Pantano ◽  
...  
Keyword(s):  
Pilot Study ◽  
Oral Fluid ◽  
Urine Samples ◽  
Screening Tests ◽  
Gas Chromatography Mass Spectrometry ◽  
Negative Results ◽  
A Value ◽  
Order Of Magnitude ◽  
Confirmation Analysis

Abstract Background In those countries where cannabis use is still illegal, some manufacturers started producing and selling “light cannabis”: dried flowering tops containing the psychoactive principle Δ-9-tetrahydrocannabinol (THC) at concentrations lower than 0.2% together with variable concentration of cannabidiol (CBD). We here report a pilot study on the determination of cannabinoids in the oral fluid and urine of six individuals after smoking 1 g of “light cannabis”. Methods On site screening for oral fluid samples was performed, as a laboratory immunoassay test for urine samples. A validated gas chromatography-mass spectrometry (GC-MS) method was then applied to quantify THC and CBD, independently from results of screening tests. Results On site screening for oral fluid samples, with a THC cut-off of 25 ng/mL gave negative results for all the individuals at different times after smoking. Similarly, negative results for urine samples screening from all the individuals were obtained. Confirmation analyses showed that oral fluid THC was in the concentration range from 2.5 to 21.5 ng/mL in the first 30 min after smoking and then values slowly decreased. CBD values were usually one order of magnitude higher than those of THC. THC-COOH, the principal urinary THC metabolite, presented the maximum urinary value of 1.8 ng/mL, while urinary CBD had a value of 15.1 ng/mL. Conclusions Consumers of a single 1 g dose of “light cannabis” did not result as positive in urine screening, assessing recent consumption, so that confirmation would not be required. Conversely, they might result as positive to oral fluid testing with some on-site kits, with THC cut-off lower than 25 ng/mL, at least in the first hour after smoking and hence confirmation analysis can be then required. No conclusions can be drawn of eventual chronic users.

scholarly journals Essais interlaboratoires pour l'identification moléculaire des espèces paléarctiques du sous-genre Avaritia

2009 ◽  
Vol 62 (2-4) ◽  
pp. 102
Author(s):  
Catherine Cetre-Sossah ◽  
Thomas Balenghien ◽  
Jean-Claude Delécolle ◽  
Rudolf Meiswinkel
Keyword(s):  
Deoxyribonucleic Acid ◽  
False Negative ◽  
Chain Reaction ◽  
Negative Results ◽  
False Negative Results ◽  
Ring Trial ◽  
International Experts ◽  
Phosphate Buffered Saline ◽  
Excel File

A ring trial was conducted for molecular identification of Palaearctic species of the subgenus Avaritia, and especially the following four species: Culicoides chiopterus, C. dewulfi, C. obsoletus and C. scoticus. It was based on multiplex polymer­ase chain reaction (PCR) on the molecular markers cytochrome oxidase type 1 (CO1), internal transcribed spacer 2 (ITS-2), and ITS-1. Each of the 13 participating laboratories (from seven dif­ferent countries) received on the 4th of August 2008 a panel of 38 samples of 11 μL of a phosphate-buffered saline (PBS) solu­tion containing parts of a single specimen of insect ground up into 200 μL of PBS, as well as four tubes, identified and men­tioned in the accompanying letter, for which deoxyribonucleic acid (DNA) had already been extracted.  The panel was coded with a letter followed by different num­bers. The laboratories had two months from the date of arrival of the samples to give back the results by sending an Excel file containing the coding. The 38 samples used for the trial were exchanged for identification between two international experts (Drs J.C. Delécolle and R. Meiswinkel). Only one identification differed between the two experts: scoticus vs. obsoletus, and sequencing revealed it to be a C. scoticus specimen.  Only one laboratory used molecular marker ITS-2, five labo­ratories used ITS-1, and four used CO1. Only two found the expected results. The eight remaining laboratories found some false positive or false negative results. Five out of ten correctly identified the species from the DNA samples. Seven out of ten laboratories had 100% sensitivity

Competitive Direct Enzyme-Linked Immunosorbent Assay for Detection of Sulfamethazine Residues in Swine Urine and Muscle Tissue

1988 ◽  
Vol 71 (6) ◽  
pp. 1137-1140 ◽  
Author(s):  
Deborah E Dixon-Holland ◽  
Stanley E Katz
Keyword(s):  
Horseradish Peroxidase ◽  
Muscle Tissue ◽  
Immunosorbent Assay ◽  
Microtiter Plate ◽  
Test System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sensitive Assay ◽  
Saline Extract ◽  
Swine Urine ◽  
Phosphate Buffered Saline

Abstract A sensitive assay for the detection of sulfamethazine in swine urine and muscle tissue using a direct competitive enzyme-linked immunosorbent assay (ELISA) has been developed. Undiluted urine or a phosphate-buffered saline extract of pork muscle tissue is mixed with an enzyme-labeled conjugate of sulfamethazine and horseradish peroxidase. The mixture is added to wells of a microtiter plate coated with antibody to sulfamethazine. After the test system is incubated, washed, and re-incubated with substrate and the reaction is stopped, the absorbance is measured at 405 nm. Levels of sulfamethazine as low as 20 ng sulfamethazine/g muscle tissue and 10 ng sulfamethazine/ mL swine urine were detected and estimated

scholarly journals Excretion and Detection of Cathinone, Cathine, and Phenylpropanolamine in Urine after Kath Chewing

2002 ◽  
Vol 48 (10) ◽  
pp. 1715-1719 ◽  
Author(s):  
Stefan W Toennes ◽  
Gerold F Kauert
Keyword(s):  
Analytical Techniques ◽  
Urine Samples ◽  
Gas Chromatography Mass Spectrometry ◽  
Fluorescence Polarization Immunoassay ◽  
Photodiode Array Detection ◽  
Negative Results ◽  
Specificity And Sensitivity ◽  
Photodiode Array ◽  
Array Detection ◽  
Positive Results

Abstract Introduction: The stimulating herbal drug kath is uncommon in most countries, and information on its detection and interpretation of analytical results is limited. Therefore, a study with kath was carried out to compare the efficiencies of different analytical techniques used to detect drug use. Methods: Four volunteers chewed kath leaves for 1 h; urine samples were collected up to 80 h afterward and analyzed by the Abbott fluorescence polarization immunoassay (FPIA), the Mahsan-AMP300 on-site immunoassay, the Bio-Rad Remedi HS HPLC system with photodiode array detection (DAD), and gas chromatography–mass spectrometry (GC-MS). Results: FPIA gave negative results, whereas positive results were obtained with the Mahsan test during the first day. With HPLC, one peak could be observed up to 50 h, but its DAD spectrum could not be identified by the system. Further investigations indicated that the kath alkaloids coeluted and produced a mixed DAD spectrum. With GC-MS, the specific kath ingredient cathinone was detected up to 26 h, whereas cathine and norephedrine were still detectable in the last samples. Maximum concentrations of cathinone, cathine, and norephedrine in urine samples from the study were 2.5, 20, and 30 mg/L, respectively, whereas in authentic cases the concentrations were much higher. Conclusion: GC-MS is superior to the screening techniques Mahsan-AMP300 and Remedi with respect to specificity and sensitivity for the detection of kath use in urine.

Measuring Urinary Glycosaminoglycans in the Presence of Protein: An Improved Screening Procedure for Mucopolysaccharidoses Based on Dimethylmethylene Blue

1992 ◽  
Vol 38 (6) ◽  
pp. 803-807 ◽  
Author(s):  
J G de Jong ◽  
R A Wevers ◽  
R Liebrand-van Sambeek
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
False Negative ◽  
Urine Samples ◽  
Screening Procedure ◽  
Urinary Proteins ◽  
Negative Results ◽  
False Negative Results ◽  
Metabolic Screening ◽  
Urinary Glycosaminoglycans

Abstract Earlier we described a simple and reliable screening procedure in urine for mucopolysaccharidoses based on the color reaction of glycosaminoglycans (GAGs) with dimethylmethylene blue. At physiological concentrations of urinary protein, we observed an obvious interference by protein in the assay. By modifying the assay, we abolished the protein interference. The modified procedure is not disturbed by human serum albumin, IgG (both tested with as much as 5 g/L of protein), or urinary proteins. The modified procedure appeared as reliable as the original. No false-negative results were found in a series of 26 urine samples from patients with mucopolysaccharidoses (sensitivity 100%). In a series of 405 urine samples offered for metabolic screening, 24 samples with increased GAG content and normal GAG composition were seen (specificity 94%). The method may also be applicable for determining GAG in other body fluids or solutions containing protein.

Cloning and sequencing of the ompL37 gene present in Leptospira interrogans, a surface protein in pathogenic leptospires

2019 ◽  
Author(s):  
Elaheh Rezaei ◽  
Pejvak Khaki ◽  
Soheila Moradi Bidhendi ◽  
Mojtaba Noofeli ◽  
Maryam Sadat Soltani
Keyword(s):  
Binding Sites ◽  
Diagnostic Tests ◽  
Genomic Dna ◽  
Surface Protein ◽  
Primary Objective ◽  
Leptospira Interrogans ◽  
Cloning And Sequencing ◽  
Future Studies ◽  
Conserved Sequence ◽  
Pcr Products

Background and Objectives: Leptospirosis, an infection caused by pathogenic leptospires, is associated with insufficient sanitation and poverty. Leptospira is transmitted through contact with contaminated urine of reservoir animals. The primary objective of this study was to clone and sequence the ompL37 gene present in local and vaccine serovars. Materials and Methods: A total of 16 Leptospira interrogans serovars were cultured in EMJH liquid medium. After grow- ing, genomic DNA was extracted using phenol-chloroform method. Primer pair was synthesized to amplify the 996 bp ompL37 sequence. The amplified ompL37 gene was cloned into pTZ57R/T vector. The sequences obtained from this study were compared with an only recorded sequence in the Genbank by the Meg Align software. Results: PCR products showed an amplified 996bp ompL37 gene product belonging to pathogenic serovars, while no ompL37 products were amplified in non-pathogenic serovars. Sequences comparison tests from 16 native serotypes exam- ined in this study displayed a similarity range of 84% to 99.5% among serovars used. The results showed that two serotypes of L. interrogans including Serjoehardjo (RTCC2810 and RTCC2821) had the highest identity up to 95.5%. Two serovars of L. interrogans including Pomona (RTCC2822) and Icterohaemorrhagiae (RTCC2823) had the lowest identity about 84%. Conclusion: As the results showed, ompL37, present on the surface of such bacteria, showed a conserved sequence. ompL37, as a key role in cell adhesion and pathogenicity, can be used for designing diagnostic tests and vaccines. Furthermore, se- quencing of various sites in ompL37 gene, including binding sites and immunogenic epitopes, can be valuable alternatives for future studies.

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