Molecular diagnostic investigation of brucellosis in a caprine organic farm

Brucellosis of sheep and goats is widely spread in the Mediterranean basin. The disease is of considerable significance with connection to Public Health protection since it can be transmitted to humans causing serious disease. In April 2005 we investigated whether brucellosis was present among the male animals of a caprine organic herd in Nomos Hleias, Greece. The herd consisted of 250 female and 13 male animals and had a record of sporadic abortions usually taking place at the final third of gestation. During sample collection all the animals were found clinically healthy, although it was not possible to determine from the records of the farm if there was any previous incidence of orchitis or epididymitis in the animals under study. The laboratory diagnostic investigation consisted of a polymerase chain reaction (PCR) assay applied for the detection of DNA belonging to Brucella melitensis in blood samples. Two of the 13 samples that were tested reacted positive by PCR allowing the amplification of the 844 base pair DNA fragment specific for Brucella melitensis. In this study, PCR allowed detection of two animals that although they were clinically healthy, they carried enough Brucella in their blood to allow microbial identification with only a single blood collection. This finding seems to agree with the concept that goats consist the reservoir of brucellosis in Greece developing milder disease than sheep and sustaining the infection for longer periods of time. One of the positive samples that were recorded was identified by sequencing as Brucella suis, something that is reported for the first time with connection to goats. This finding and the sensitivity of man to B. suis renders human exposure to this pathogen through goats, an epidemiology factor worth of detailed investigation. This necessity is associated with the fact that as opposed to porcine meat, sheep and goat dairy products in Greece are sometimes consumed without the necessary heat treatment.

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Clinical Evaluation of the Torq Zero Delay Centrifuge System for Decentralized Blood Collection and Stabilization

Diagnostics ◽

10.3390/diagnostics11061019 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1019

Author(s):

Kyungjin Hong ◽

Gabriella Iacovetti ◽

Ali Rahimian ◽

Sean Hong ◽

Jon Epperson ◽

...

Keyword(s):

Clinical Chemistry ◽

Blood Collection ◽

Test Results ◽

Sample Collection ◽

Rapid Separation ◽

Cell Counts ◽

Collection Device ◽

Precision Study ◽

Clinically Significant ◽

Blood Specimens

Blood sample collection and rapid separation—critical preanalytical steps in clinical chemistry—can be challenging in decentralized collection settings. To address this gap, the Torq™ zero delay centrifuge system includes a lightweight, hand-portable centrifuge (ZDrive™) and a disc-shaped blood collection device (ZDisc™) enabling immediate sample centrifugation at the point of collection. Here, we report results from clinical validation studies comparing performance of the Torq System with a conventional plasma separation tube (PST). Blood specimens from 134 subjects were collected and processed across three independent sites to compare ZDisc and PST performance in the assessment of 14 analytes (K, Na, Cl, Ca, BUN, creatinine, AST, ALT, ALP, total bilirubin, albumin, total protein, cholesterol, and triglycerides). A 31-subject precision study was performed to evaluate reproducibility of plasma test results from ZDiscs, and plasma quality was assessed by measuring hemolysis and blood cells from 10 subject specimens. The ZDisc successfully collected and processed samples from 134 subjects. ZDisc results agreed with reference PSTs for all 14 analytes with mean % biases well below clinically significant levels. Results were reproducible across different operators and ZDisc production lots, and plasma blood cell counts and hemolysis levels fell well below clinical acceptance thresholds. ZDiscs produce plasma samples equivalent to reference PSTs. Results support the suitability of the Torq System for remotely collecting and processing blood samples in decentralized settings.

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To Determine The Frequency Of Vitamin D Deficiency In Patients With Chronic Hepatitis C

Journal of Bahria University Medical and Dental College ◽

10.51985/jbumdc2018065 ◽

2018 ◽

Vol 08 (04) ◽

pp. 241-244

Author(s):

Imran Hussain ◽

M. Zill-e-Humayun Mirza ◽

Ali Yusuf

Keyword(s):

Vitamin D ◽

Chronic Hepatitis ◽

Hepatitis C ◽

Vitamin D Deficiency ◽

Chronic Hepatitis C ◽

Cross Sectional Study ◽

Blood Collection ◽

Study Cohort ◽

Sample Collection ◽

Cross Sectional

Objective: To determine the frequency of vitamin D deficiency in patients with chronic hepatitis C (CHC) Design: It was a Descriptive and Cross Sectional study Place and Duration of Study: It was carried out in the Medicine Unit of Pakistan Naval Ship SHIFA, Karachi from Nov 29, 2016 to May 29, 2017. Patients and Methods: Approval was sought from Institutional Review Board before carrying out the study. Proper history, clinical examination and appropriate lab investigations were carried out. Standard techniques were used for blood sample collection. Site used for blood collection was antecubital fossa. Sterile method was used for fasting sample and about 10 ml of blood was collected from each patient. Results: A total of 289 patients were included. Strict exclusion and inclusion criteria was used for study cohort. Mean age (years) of study cohort was 34.51+8.32. There were 188 (65.1) male and 101 (34.9) female patients. Patients with CHC who were vitamin D deficient were 74 (25.6). Conclusion: Patients of CHC had high frequency of vitamin D deficiency which suggests that further studies in the region will be conduct in our general population to know the exact statistics which will pave the way for future researchers

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Potential natural enemies of the invasive rugose spiraling whitefly, Aleurodicus rugioperculatus Martin in India

Journal of Biological Control ◽

10.18311/jbc/0/15598 ◽

2017 ◽

Vol 30 (4) ◽

pp. 236 ◽

Cited By ~ 2

Author(s):

K. Selvaraj ◽

R. Sundararaj ◽

T. Venkatesan ◽

Chandish R. Ballal ◽

S. K. Jalali ◽

...

Keyword(s):

Natural Enemies ◽

Tamil Nadu ◽

Ornamental Plants ◽

Coi Gene ◽

Molecular Diagnostic ◽

Pest Species ◽

Molecular Tools ◽

Accession Number ◽

Custard Apple ◽

First Time

A invasive rugose spiraling whitefly (RSW) Aleurodicus rugioperculatus Martin (Hemiptera: Aleyrodidae) was found infesting coconut, banana, custard apple and several ornamental plants in Tamil Nadu, Andhra Pradesh and Kerala for the first time in India. The identity of the pest species was determined through morphological and molecular tools. Furthermore cytochrome c oxidase-I gene (658 bp) of RSW was sequenced (GenBank accession number KY209909) which would serve as an ideal molecular diagnostic marker for its identification irrespective of its phenotypic plasticity. During the survey, several natural enemies were recorded and maximum parasitism was recorded by Encarsia guadeloupae Viggiani (Hymenoptera: Aphelinidae) and its COI gene was sequenced and deposited as Encarsia sp. (GenBank accession number KY223606). Per cent parasitism ranged from 20.0 to 60.0 % in different collection locations, highest parasitism being recorded in Kerala as compared to other states. The predators recorded were Mallada sp., few coccinellids and predatory mites. This communication is the first report of the rugose spiraling whitefly, its host plant range and associated natural enemies in India.

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Frequency of molecular detection of equine coronavirus in faeces and nasal secretions in 277 horses with acute onset of fever

Veterinary Record ◽

10.1136/vr.104919 ◽

2019 ◽

Vol 184 (12) ◽

pp. 385-385 ◽

Cited By ~ 1

Author(s):

Nicola Pusterla ◽

Kaitlyn James ◽

Samantha Mapes ◽

Farifield Bain

Keyword(s):

Clinical Signs ◽

Acute Onset ◽

Molecular Diagnostic ◽

Nasal Discharge ◽

Respiratory Pathogens ◽

Sample Collection ◽

Diagnostic Laboratory ◽

Study Results ◽

Streptococcus Equi ◽

Nasal Secretions

ContextDue to the inconsistent development of enteric signs associated with ECoV infection in adult horses, many practitioners collect nasal secretions rather than feces for the molecular diagnostic work-up of such horses.Main conclusionECoV infection should be considered in horses presenting with acute onset of fever, especially when nasal discharge is absent as one of the cardinal clinical sign.ApproachA total of 277 adult horses with acute onset of fever were enrolled in this study. Feces were tested for ECoV and nasal secretions for common respiratory pathogens (equine herpesvirus (EHV)-1, EHV-4, equine influenza virus (EIV), equine rhinitis viruses (ERVs) and Streptococcus equi ss. equi) and ECoV by qPCR. Each submission was accompanied by a questionnaire requesting information pertaining to signalment, use, recent transportation, number of affected horses on the premise and presence of clinical signs at the time of sample collection.ResultsThe total number of horses testing qPCR-positive for ECoV in feces was 20 (7.2%), 4 of which also tested qPCR-positive for ECoV in nasal secretions. In the same population 9.0% of horses tested qPCR-positive for EHV-4, 6.1% for EIV, 4.3% for Streptococcus equi ss. equi, 3.2% for ERVs and 0.7% for EHV-1. Draft horses, pleasure use, multiple horses affected on a premise and lack of nasal discharge were significantly associated with ECoV qPCR-positive horses.InterpretationThe present study results showed that 7.2% of horses with acute onset of fever tested qPCR-positive for ECoV in feces, highlighting the importance of testing such horses for ECoV in feces. The various prevalence factors associated with ECoV qPCR-positive status likely relate to the high infectious nature of ECoV and breed-specific differences in management and husbandry practices.Significance of findingsECoV infection should be suspected and tested for in horses presenting with acute onset of fever, lethargy and anorexia with no respiratory signs. A two-step approach should be consider in which respiratory secretions and feces should be collected from such horses and submitted to a diagnostic laboratory. If the respiratory secretions test negative by qPCR for a panel of respiratory pathogens, feces already submitted to the laboratory should be tested for ECoV.

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The integrated blood-collection system as a vehicle into complete clinical laboratory automation

Clinical Chemistry ◽

10.1093/clinchem/37.9.1548 ◽

1991 ◽

Vol 37 (9) ◽

pp. 1548-1556 ◽

Cited By ~ 8

Author(s):

R L Columbus ◽

H J Palmer

Keyword(s):

Clinical Laboratory ◽

Integrated System ◽

Diagnostic Instrument ◽

Laboratory Evaluation ◽

Blood Collection ◽

Sample Collection ◽

Collection System ◽

Patient Identification ◽

Plasma Extraction ◽

Derivatives Of

Abstract A rationale is offered and methodology illustrated for integrating the fundamental steps involved in the collection and processing of blood for laboratory evaluation. The approach taken in the development of these concepts and components greatly extends the possibilities of laboratory systems integration without upsetting established modalities. A prototype design of the integrated blood-collection system integrates blood collection, cellular separations, sample transfer to stable storage without chemical mediators, and sample presentation for chemical analysis (e.g., precision metering) while preserving patient identification. A sophisticated, multi-chambered blood-collection container is the site of all blood sample processing and transfer steps. This device is supported by a compact, robotic centrifuge of unique design and a transfer mechanism to facilitate sample delivery for analysis within a diagnostic instrument. The confluence of these individual components into a single integrated system provides the means to completely automate the processing of blood samples, after sample collection, eliminating all manual transfer steps and any external exposure of blood interfaces outside the diagnostic instrument. Configurational derivatives of the Integrated Blood-Collection System offer choice of skin or venipuncture procedure, rapid plasma extraction for micro- or macro-collected volumes, and sample delivery by either aspiration or direct metering of discrete 10-microL samples from the collection container. The skin-puncture configuration provides the opportunity within a single device to collect and process up to 500 microL of sample by capillarity from a skin prick.

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Maintenance of an Old World Betasatellite by a New World Helper Begomovirus and Possible Rapid Adaptation of the Betasatellite

Journal of Virology ◽

10.1128/jvi.00795-09 ◽

2009 ◽

Vol 83 (18) ◽

pp. 9347-9355 ◽

Cited By ~ 64

Author(s):

Muhammad Shah Nawaz-ul-Rehman ◽

Shahid Mansoor ◽

Rob W. Briddon ◽

Claude M. Fauquet

Keyword(s):

New World ◽

Cotton Leaf ◽

Old World ◽

Tropical Regions ◽

Cabbage Leaf Curl Virus ◽

Chili Leaf Curl Betasatellite ◽

Serious Disease ◽

Leaf Curl Virus ◽

First Time ◽

Leaf Curl

ABSTRACT Begomoviruses (family Geminiviridae) cause major losses to crops throughout the tropical regions of the world. Begomoviruses originating from the New World (NW) and the Old World (OW) are genetically distinct. Whereas the majority of OW begomoviruses have monopartite genomes and whereas most of these associate with a class of symptom-modulating satellites (known as betasatellites), the genomes of NW begomoviruses are exclusively bipartite and do not associate with satellites. Here, we show for the first time that a betasatellite (cotton leaf curl Multan betasatellite [CLCuMuB]) associated with a serious disease of cotton across southern Asia is capable of interacting with a NW begomovirus. In the presence of CLCuMuB, the symptoms of the NW cabbage leaf curl virus (CbLCuV) are enhanced in Nicotiana benthamiana. However, CbLCuV was unable to interact with a second betasatellite, chili leaf curl betasatellite. Although CbLCuV can transreplicate CLCuMuB, satellite accumulation levels in plants were low. However, progeny CLCuMuB isolated after just one round of infection with CbLCuV contained numerous mutations. Reinoculation of one such progeny CLCuMuB with CbLCuV to N. benthamiana yielded infections with significantly higher satellite DNA levels. This suggests that betasatellites can rapidly adapt for efficient transreplication by a new helper begomovirus, including begomoviruses originating from the NW. Although the precise mechanism of transreplication of betasatellites by begomoviruses remains unknown, an analysis of betasatellite mutants suggests that the sequence(s) required for maintenance of CLCuMuB by one of its cognate begomoviruses (cotton leaf curl Rajasthan virus) differs from the sequences required for maintenance by CbLCuV. The significance of these findings and, particularly, the threat that betasatellites pose to agriculture in the NW, are discussed.

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Porcine reproductive and respiratory syndrome virus RNA detection in different matrices under typical storage conditions in the UK

Veterinary Record ◽

10.1136/vr.105312 ◽

2019 ◽

Vol 185 (1) ◽

pp. 21-21 ◽

Cited By ~ 1

Author(s):

Jinghui Fan ◽

Priscilla F Gerber ◽

Ana Cubas Atienzar ◽

Lysan Eppink ◽

Chong Wang ◽

...

Keyword(s):

Storage Condition ◽

Early Morning ◽

Respiratory Syndrome Virus ◽

Blood Collection ◽

Practical Reasons ◽

Sample Type ◽

Sample Collection ◽

Blood Samples ◽

Fta Cards ◽

The Uk

In the UK, approximately 40 per cent of the pig breeding herds are outdoors. To monitor their porcine reproductive and respiratory syndrome virus (PRRSV) status, blood is collected commonly from piglets around weaning. Sample collection in British outdoor pigs often occurs during the early morning hours when the piglets tend to accumulate inside sheltered areas. For practical reasons, dry cotton swabs are occasionally used for blood collection and stored at room temperature until arrival in the laboratory. Detection of PRRSV RNA is a function of viral concentration, sample type and storage condition. To evaluate a possible impact of the sampling protocol on PRRSV1 detection, experimentally spiked blood samples using three dilutions of a representative PRRSV1 strain were prepared. In addition, blood samples from pigs naturally infected with PRRSV were obtained from a PRRSV-positive British herd. Spiked blood and blood from infected pigs were used to obtain sera, dry or wet (immersed in saline) polyester or cotton swabs and FTA cards. The different samples were stored for 24 hours, 48 hours or 7 days at 4°C or 20°C and tested by a real-time reverse transcriptase PRRSV PCR assay. Under the study conditions, the best matrix was serum (96.7 per cent), followed by wet swabs (78 per cent), dry swabs (61.3 per cent) and FTA cards (51 per cent). Polyester swabs (76 per cent) showed a better performance than cotton swabs (63.3 per cent). The reduction in sensitivity obtained for swabs and FTA cards was particularly high at low viral concentrations. The results indicate that wet polyester swabs should be used whenever possible.

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Challenges for conducting blood collection and biochemical analysis in a large multicenter school-based study with adolescents: lessons from ERICA in Brazil

Cadernos de Saúde Pública ◽

10.1590/0102-311x00122816 ◽

2017 ◽

Vol 33 (4) ◽

Cited By ~ 9

Author(s):

Felipe Vogt Cureau ◽

Katia Vergetti Bloch ◽

Aline Henz ◽

Camila W. Schaan ◽

Carlos Henrique Klein ◽

...

Keyword(s):

Cardiovascular Risk ◽

Air Transportation ◽

Blood Collection ◽

Reference Laboratory ◽

Sample Collection ◽

Temperature Changes ◽

School Based ◽

Methodological Aspects ◽

Single Laboratory ◽

Metabolic Syndrome Components

Abstract: The Study of Cardiovascular Risk in Adolescents (ERICA) is a pioneering study that aimed to assess the prevalence of cardiovascular risk factors, including metabolic syndrome components in Brazilian adolescents. This study aims to describe the methodological aspects related to blood collection as well as to report pertaining results of the preparation, transport, storage, and exams in ERICA. Exams in ERICA were performed in a single laboratory and blood samples were collected in schools in a standardized manner. Logistics involved air transportation of samples to the reference laboratory with controlled temperature since sample collection. The serum was stored in local biorepositories in four centers to be used in future analyses. During the study, 284,247 exams were performed and rate of participation in exams was 56.2%, thus involving 40,732 adolescents. From the total, 92.6% of the samples reached the reference laboratory maintaining the temperature between 0-10°C. No clinical significant changes in results due to temperature changes were identified. External quality control recorded satisfactory results in 98.7% of the evaluations. Four biorepositories with samples of 7,785 adolescents were created. Thus, we can consider that the logistics adopted in ERICA was fairly successful and description of this as well as the difficulties experienced in Brazil can inform and facilitate the planning of future studies, especially in developing countries.

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The influence of different sample collection types on the levels of markers used for Down's syndrome screening as measured by the Kryptor Immunosassay system

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1258/000456303763046102 ◽

2003 ◽

Vol 40 (2) ◽

pp. 166-168 ◽

Cited By ~ 22

Author(s):

Kevin Spencer

Keyword(s):

Processing Time ◽

Point Of Care ◽

Protein A ◽

First Trimester ◽

Maternal Serum ◽

Blood Collection ◽

Chromosomal Anomalies ◽

Sample Collection ◽

Edta Plasma ◽

Β Hcg

Background: In a rapid point-of-care screening programme for chromosomal anomalies, analysis of biochemical markers in maternal blood can now be accomplished in a rapid time frame (less than 20 min). The need to leave whole blood samples some 10 min for coagulation and a further 5 min for centrifugation adds additional processing time. Methods: The possibilities for reducing this processing time were investigated using various anticoagulated blood collection systems and the Kryptor analytical platform. Plasma levels of α-fetoprotein (AFP), pregnancy-associated plasma protein-A (PAPP-A) and free human chronic gonadotrophin β-subunit (β-hCG) were compared with those in maternal serum. Results: From the mean results from ten patients it was shown that use of heparin plasma resulted in a statistically significant reduction in levels of PAPP-A and that EDTA plasma reduced the levels of PAPP-A dramatically. For AFP, levels in citrated plasma and EDTA plasma were also significantly reduced, whereas levels of free β-hCG were not affected. Conclusion: Use of alternative sample types for PAPP-A is not possible. The sample of choice for first trimester screening using the Kryptor platform is maternal serum.

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Extraction-free rapid cycle RT-qPCR and extreme RT-PCR for SARS-CoV-2 virus detection

American Journal of Clinical Pathology ◽

10.1093/ajcp/aqab191.296 ◽

2021 ◽

Vol 156 (Supplement_1) ◽

pp. S139-S139

Author(s):

J C Lownik ◽

J S Farrar ◽

G Way ◽

R K Martin

Keyword(s):

Supply Chain ◽

Diagnostic Testing ◽

Rna Extraction ◽

Virus Detection ◽

Detection Methods ◽

Molecular Diagnostic ◽

Sample Collection ◽

Rt Pcr ◽

Hydrolysis Probe ◽

Time Requirements

Abstract Introduction/Objective Since the start of the coronavirus disease 2019 (COVID-19) pandemic, molecular diagnostic testing for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has faced substantial supply chain shortages and noteworthy delays in result reporting after sample collection. Supply chain shortages have been most evident in reagents for RNA extraction and rapid diagnostic testing. In this study, we explored the kinetic limitations of extraction-free rapid cycle RT-qPCR for SARS-CoV-2 virus detection using the commercially available capillary based LightCycler. Methods/Case Report We optimized reverse transcription and PCR under extraction-free and rapid thermocycling conditions utilizing hydrolysis probe-based detection methods using a Roche LightCycler. Results (if a Case Study enter NA) This protocol improves detection speed while maintaining the sensitivity and specificity of hydrolysis probe-based detection. Percentage agreement between the developed assay and previously tested positive patient samples was 97.6% (n= 40/41) and negative patient samples was 100% (40/40). We further demonstrate that using purified RNA, SARS-CoV-2 testing using extreme RT-PCR and product verification by melting can be completed in less than 3 minutes. Conclusion We developed a protocol for sensitive and specific RT-qPCR of SARS-CoV-2 RNA from nasopharyngeal swabs in less than 20 minutes, with minimal hands-on time requirements. Overall, these studies provide a framework for increasing the speed of SARS-CoV-2 and other infectious disease testing.

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