scholarly journals The Use of Multiplex Phosphorescence Analysis (PHOSPHANTM) and Polymerase Chain Reaction for Laboratory Diagnosis of Ixodid Tick-borne Borrelioses

2015 ◽  
Vol 14 (2) ◽  
pp. 38-44
Author(s):  
T. I. Kuznetsova ◽  
V. G. Pomelova ◽  
E. I. Korenberg ◽  
N. S. Osin
Keyword(s):  
Polymerase Chain Reaction ◽  
High Efficiency ◽  
Erythema Migrans ◽  
Clinical Signs ◽  
Laboratory Diagnosis ◽  
Ixodid Tick ◽  
Chain Reaction ◽  
Cutaneous Manifestations ◽  
Granulocytic Anaplasmosis ◽  
Polymerase Chain

In this report, we evaluated the performance of C6 peptide based multiplex Phosphorescence Analysis (PHOSPHANTM) and Polymerase Chain Reaction (nested PCR) for laboratory diagnosis of Ixodid Tick-borne Borrelioses (ITBB). The study was conducted on 155 patients with localized and disseminated stages of the disease, the cases of mixed infection with ITBB and human granulocytic anaplasmosis including. Positive PHOSPHAN reactions were observed in 78 ± 7.7% of patients with erythema migrans (EM) and 91 ± 11.7% of patients without cutaneous manifestations of the disease. The frequency of PCR positive samples was lower, 26 ± 8.2% and 72 ± 17.1% respectively. The maximum frequency of positive samples detected by both methods was mainly observed at 2 - 4 week from the onset of the disease (or 22 - 35 day after tick bite). In general, PHOSPHAN provided serologic confirmation of the disease in 52 of 55 (94.5 ± 6.2%) patients, whose blood contained Borrelia DNA. Only 3 patients tested positive in PCR (1 - with EM and 2 - without this skin manifestation) were seronegative. These data confirmed the high efficiency of PHOSPHAN method for serologic verification of ITBB both at localized and disseminated stages of the disease. The use of PCR (in addition to PHOSPHAN) is appropriate within a certain period of time (no later than 2 - 3 weeks from the onset of the disease) to clarify the diagnosis in seronegative patients having clinical signs of disseminated non-cutaneous form of ITBB, or atypical cutaneous manifestations of erythematous form of the disease.

scholarly journals Comparison of Three Rapid Commercial Canine Parvovirus Antigen Detection Tests with Electron Microscopy and Polymerase Chain Reaction

2009 ◽  
Vol 21 (3) ◽  
pp. 344-345 ◽  
Author(s):  
Silke Schmitz ◽  
Christina Coenen ◽  
König Matthias ◽  
Thiel Heinz-Jürgen ◽  
Reto Neiger
Keyword(s):  
Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Chronic Diarrhea ◽  
Gastrointestinal Disease ◽  
Clinical Signs ◽  
Canine Parvovirus ◽  
Chain Reaction ◽  
Group A ◽  
Polymerase Chain ◽  
Group B

Different antibody-based tests for rapid detection of Canine parvovirus antigens in feces are commercially available, allowing quick diagnosis in a clinical setting. However, the diagnostic accuracy of these tests compared with standard methods has not been evaluated so far. In the current study, 3 commercial tests were compared with immune-electron microscopy (IEM) and polymerase chain reaction (PCR). Dogs were divided into 3 groups: group A, samples from dogs with acute hemorrhagic diarrhea ( n = 50); group B, dogs with chronic diarrhea ( n = 10); and group C, dogs with no evidence of gastrointestinal disease ( n = 40). Specificity of all 3 commercial tests versus PCR and IEM was good to excellent (92.2–100%). Sensitivity, in contrast, was poor: 15.8–26.3% versus PCR and 50–60% versus IEM. In group A, 10 dogs were positive by IEM and 24 dogs were positive by PCR. Positive PCR results were also obtained from animals in control groups (group B, 1 dog; group C, 5 dogs). No dog in group B or C was positive by IEM. In conclusion, the rapid tests are useful to diagnose canine parvoviral enteritis, but they do not rule out parvovirus infection in an animal with typical clinical signs. In addition, a small percentage of healthy dogs and dogs with chronic diarrhea showed positive PCR results; this may be due to asymptomatic/persistent infection or intestinal passage of virus. The significance of this finding remains unclear.

scholarly journals RAPID AND SPESIFIC DETECTION OF MYCOBACTERIUM TUBERCULOSIS USING POLYMERASE CHAIN REACTION

2019 ◽  
Vol 3 (2) ◽  
pp. 83
Author(s):  
Anita Kurniati ◽  
Desak Nyoman Surya Suameitra Dewi ◽  
Ni Nyoman Purwani
Keyword(s):  
Polymerase Chain Reaction ◽  
Mycobacterium Tuberculosis ◽  
Clinical Specimen ◽  
Laboratory Diagnosis ◽  
Nucleic Acid Amplification ◽  
Valuable Insight ◽  
Chain Reaction ◽  
Severe Condition ◽  
Middle Income ◽  
Polymerase Chain

Background: Tuberculosis (TB) is one of the major causes of health burden worldwide, especially in lower middle-income countries. TB is caused by Mycobacterium tuberculosis (MTB) and characterized by severe condition incuding coughing and fever. Purpose: To review the current methods for detection of TB using Polymerase Chain Reaction (PCR). Review: several studies have been done to give valuable insight into TB transmission, diagnosis, and treatment, however research  is constantly  needed  to decrease the incidence of eradicate TB. This infectious disease still give big health problem in all over the world by being second in causing high mortality rates after HIV/AIDS.  A specific, sensitive, rapid and cheap method for TB and other mycobacteria diagnosis in clinical specimen is a desperate needed in the laboratory diagnosis and hence management of tuberculosis. PCR as one of nucleic acid amplification assays have revolutionized MTB detection. Since it was first invented in fifteen years ago, it’s been through many developments. Conclusion: PCR  is one of the most specific and sensitive method currently available for TB diagnosis that can also detect in in all types of specimens obtained from TB patients.

Thermal Analysis of a Micro-Polymerase Chain Reaction Device

2004 ◽  
Author(s):  
Jeff Punch ◽  
Bryan Rodgers ◽  
David Newport ◽  
Mark Davies
Keyword(s):  
Thermal Analysis ◽  
Polymerase Chain Reaction ◽  
High Efficiency ◽  
Closed Form Solution ◽  
Thermal Control ◽  
Rate Of Change ◽  
Temperature Cycling ◽  
Chain Reaction ◽  
Macro Scale ◽  
Polymerase Chain

Micro-scale polymerase chain reaction (micro-PCR) systems offer substantial advantages over macro-scale systems. Smaller sample volumes are required, and faster process times are feasible. Thermal control of micro-PCR systems is a substantial technical challenge, however. The PCR process requires the fluid sample to be cycled through three temperature ranges — typically 90–95°C, 50–65°C and 72–77°C for denaturation, hybridisation and replication respectively. Durations of the three steps are required to be in the ratio of 4:4:9. In this paper, the thermal analysis of a continuous flow micro-PCR device is reported. The objective of the analysis is to optimize the thermal performance of the device for fast amplification cycles with high efficiency - an efficient PCR features rapid heating and cooling between steps, and good temperature uniformity within each step. The device comprises an array of parallel microchannels formed within a polypropylene substrate to carry fluid, with the base of the substrate mounted on an aluminium carrier. Substrate depth is 500 micron, and each channel is 60 micron wide by 40 micron deep. Thermoelectric cells (TECs) are bonded to the carrier, and powered by a thermoelectric controller with feedback from sensors embedded in the carrier. A Pyrex Glass slide is bonded to the substrate to form closed channels. Arrays of film heaters mounted on the slide adjacent to the channel are used to establish the required temperature regions along the channel. By pumping the fluid at a fixed flow rate, temperature cycling of specific period is achieved. Thermal analysis of the substrate is performed using an approximate closed-form solution, in conjunction with Finite Element (FE) and Computational Fluid Dynamics (CFD) simulations. The analysis is used to conduct a parametric study in order to determine the optimum configurations of substrate materials, cooling conditions, heaters and flow rates required to impose specific temperature cycles. The use of thermoelectric cells is shown to increase the rate of change of temperature between the various regions, improving the efficiency and decreasing the cycle time of the PCR process. Cycle times of 6s or less are shown to be feasible, yielding benefits in time saved for multiple amplifications. Finally, the analysis is also used to identify the dimensionless parameters which govern the thermal characteristics of the device, illustrating the importance of the Biot number.

Systemic Mycobacterium avium Infection in a Dog Diagnosed by Polymerase Chain Reaction Analysis of Buffy Coat

2005 ◽  
Vol 41 (2) ◽  
pp. 128-132 ◽  
Author(s):  
James F. Naughton ◽  
Katrina L. Mealey ◽  
K. Jane Wardrop ◽  
J. Lindsay Oaks ◽  
Daniel S. Bradway
Keyword(s):  
Polymerase Chain Reaction ◽  
Clinical Signs ◽  
Mycobacterial Infection ◽  
Polymerase Chain Reaction Analysis ◽  
Buffy Coat ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
The Public ◽  
Avium Infection ◽  
Polymerase Chain

Dogs may be infected by Mycobacterium (M.) tuberculosis, M. bovis, and M. avium complex, and the clinical signs associated with each of these infections may be indistinguishable. Rapid speciation of the infecting organism is desirable because of the public health concerns associated with M. bovis and M. tuberculosis infections. A mycobacterial infection was suspected in the dog of this report based on acid-fast staining of organisms in macrophages obtained from liver aspirates and buffy-coat preparations. Polymerase chain reaction (PCR) analysis of a buffy-coat preparation identified M. avium.

scholarly journals Evaluation of the Reverse Transcription-Polymerase Chain Reaction/Probe Test of Serum Samples and Immunohistochemistry of Skin Sections for Detection of Acute Bovine Viral Diarrhea Infections

2002 ◽  
Vol 14 (4) ◽  
pp. 303-307 ◽  
Author(s):  
Julia F. Ridpath ◽  
Sharon K. Hietala ◽  
Steve Sorden ◽  
John D. Neill
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Amplification ◽  
Clinical Signs ◽  
Bovine Viral Diarrhea ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Serum Samples ◽  
Viral Diarrhea ◽  
Persistent Infections ◽  
Polymerase Chain

Bovine viral diarrhea viruses (BVDV) cause both acute and persistent infections. While diagnostic tests have been designed to detect animals persistently infected (PI) with BVDV, the reliability of these tests in detecting acute BVDV infections is not known. It is also possible that acute BVDV infections may be confused with persistent infections in surveys for PI animals. In this study, 2 tests presently in use in diagnostic laboratories to test for PI animals, polymerase chain reaction amplification followed by probe hybridization (RT-PCR/probe) of serum samples and immunohistochemical detection of viral antigen in skin biopsies (IHC), were evaluated for their ability to detect acute BVDV infections. Sixteen colostrum-deprived, BVDV-free, and BVDV-antibody-free calves were infected with 6 different BVDV strains. Clinical signs, seroconversion, and virus isolation indicated that inoculated animals did replicate virus. Virus could be detected in 19% (3/16) of acutely infected animals by the RT-PCR/probe technique. No acutely infected animals were positive by IHC.

scholarly journals Detection of Human Granulocytic Ehrlichiosis Agent and Borrelia Burgdorferi in Ticks by Polymerase Chain Reaction

1998 ◽  
Vol 10 (1) ◽  
pp. 56-59 ◽  
Author(s):  
Yung-Fu Chang ◽  
Vesna Novosel ◽  
Chao-Fu Chang ◽  
Jong Bae Kim ◽  
Sang J. Shin ◽  
...  
Keyword(s):  
New York ◽  
Polymerase Chain Reaction ◽  
Borrelia Burgdorferi ◽  
Laboratory Diagnosis ◽  
Etiologic Agent ◽  
Ixodid Ticks ◽  
Chain Reaction ◽  
Granulocytic Ehrlichiosis ◽  
Human Granulocytic Ehrlichiosis ◽  
Polymerase Chain

Adult ixodid ticks were collected from Westchester County, New York, and Ipswich, Massachusetts, to determine the presence of infection with a human granulocytic ehrlichiosis (HGE) agent by using the polymerase chain reaction (PCR). The presence of Borrelia burgdorferi in ticks collected from New York was also determined by PCR. Of the 229 ticks from New York and 47 ticks from Massachusetts, 9% (22/229) and 25% (12/47) of ticks contained HGE agent, respectively. Fifty-four percent (123/229) of the ticks collected from New York were B. burgdorferi positive; 4% (9/229) of these ticks contained both HGE agent and B. burgdorferi. This finding indicates that animals with Lyme borreliosis may be also exposed to the etiologic agent of HGE. More extensive laboratory diagnosis may be necessary when multiple tick-borne diseases are suspected in animals.

Evaluation of a real-time polymerase chain reaction assay for the laboratory diagnosis of giardiasis

2010 ◽  
Vol 66 (3) ◽  
pp. 261-267 ◽  
Author(s):  
Adriana Calderaro ◽  
Chiara Gorrini ◽  
Sara Montecchini ◽  
Simona Peruzzi ◽  
Giovanna Piccolo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Polymerase Chain Reaction Assay ◽  
Laboratory Diagnosis ◽  
Chain Reaction ◽  
Polymerase Chain
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