scholarly journals Molecular and serological prevalence of Toxoplasma gondii and Anaplasma spp. infection in goats from Chongqing Municipality, China

Parasite ◽  
2018 ◽  
Vol 25 ◽  
pp. 20 ◽  
Author(s):  
Zuoyong Zhou ◽  
Yutong Wu ◽  
Yiwang Chen ◽  
Zhiying Wang ◽  
Shijun Hu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Toxoplasma Gondii ◽  
Infection Rate ◽  
Anaplasma Phagocytophilum ◽  
Zoonotic Diseases ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Blood Samples ◽  
Chongqing Municipality ◽  
Polymerase Chain

Toxoplasmosis and anaplasmosis are severe zoonotic diseases, the former caused by Toxoplasma gondii and the latter by Anaplasma spp. In the present study, 332 goat blood samples were randomly collected from Chongqing Municipality, China to screen for T. gondii and Anaplasma spp. We used a polymerase chain reaction (PCR) to detect DNA, and enzyme-linked immunosorbent assay (ELISA) to test for T. gondii antibodies. The prevalence of T. gondii and Anaplasma spp. was 38% and 35% respectively by PCR, and 42% for T. gondii antibodies by ELISA. The co-infection rate by T. gondii and Anaplasma was 13%, where the two predominant pathogens co-infecting were Anaplasma phagocytophilum + A. bovis (10%), followed by T. gondii + A. phagocytophilum (9.64%). While co-infection by three pathogens varied ranging from 1.81% to 5.72%, less than 1% of goats were found to be positive for four pathogens. This is the first investigation of T. gondii and Anaplasma spp. infection in goats from Chongqing.

scholarly journals Molecular detection of Babesia bovis in cattle in Al-Qadisiyah province

2017 ◽  
Vol 40 (2) ◽  
pp. 155-158
Author(s):  
Noaman N. A,aiz
Keyword(s):  
Polymerase Chain Reaction ◽  
Infection Rate ◽  
Molecular Detection ◽  
Babesia Bovis ◽  
Chain Reaction ◽  
Adult Cattle ◽  
Blood Samples ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Age And Sex

     This study aim to determine Babesia bovis infection in cattle based on genetic methods. A total of 96 blood samples were collected from alive and slaughtered cattle from different areas in addition to the abattoir of Al-Qadisiyah province from December 2013 to August 2014. Real time polymerase chain reaction (RT.PCR) technique was used to detect the presence of the protozoan with the effect of animal's age and sex in the infection rate 47.91 % (46/96) of examined cattle were given positive result to B. bovis infection. The highest infections were shown among the adult cattle (≥1 year), while there was non-significant difference (P>0.05) in the infection rate according to the sex. So the most cattle in Al-Qadisiyah province appear to be bearing the infection predominantly as a carrier hosts.

Screening of toxoplasmosis in cancer patients: a concern

2018 ◽  
Vol 49 (1) ◽  
pp. 31-34 ◽  
Author(s):  
Amir Abdoli ◽  
Mohammad Barati ◽  
Majid Pirestani ◽  
Abdolhossein Dalimi
Keyword(s):  
Infectious Disease ◽  
Polymerase Chain Reaction ◽  
Cancer Patients ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunocompromised Patients ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
Different Types ◽  
Polymerase Chain

Toxoplasmosis is an opportunistic infectious disease in immunocompromised patients, including cancer patients, whose detection is by molecular and serological methods. A total of 106 blood samples from patients with different types of cancer were evaluated for anti- Toxoplasma gondii IgG and IgM antibodies by the enzyme-linked immunosorbent assay (ELISA) and the parasite DNA by nested polymerase chain reaction (PCR). These were detected in 41.51% (44/106) and 0.94% (1/106), respectively, but T. gondii IgM antibody was not detected at all. These results suggest that the screening of toxoplasmosis should be considered more routinely in cancer patients.

scholarly journals Use of the polymerase chain reaction to detect Toxoplasma gondii in human blood samples.

1992 ◽  
Vol 45 (10) ◽  
pp. 910-913 ◽  
Author(s):  
D O Ho-Yen ◽  
A W Joss ◽  
A H Balfour ◽  
E T Smyth ◽  
D Baird ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Toxoplasma Gondii ◽  
Human Blood ◽  
Chain Reaction ◽  
Blood Samples ◽  
Human Blood Samples ◽  
Polymerase Chain

scholarly journals Detection of Genetically Modified Corn (Bt176) in Spiked Cow Blood Samples by Polymerase Chain Reaction and Immunoassay Methods

2005 ◽  
Vol 88 (2) ◽  
pp. 654-664 ◽  
Author(s):  
Laetitia Petit ◽  
Fabienne Baraige ◽  
Yves Bertheau ◽  
Philippe Brunschwig ◽  
Annick Diolez ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Whole Blood ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Cry1ab Protein ◽  
Blood Samples ◽  
Polymerase Chain ◽  
Whole Blood Samples

Abstract The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35S promoter and Bt176-specific junction sequence) was determined by both the polymerase chain reaction–enzyme-linked immunosorbent assay (PCR–ELISA) and the 5′-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (αs1-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/mL in 10 mL whole blood samples. The sandwich ELISA kit used detected down to 1 ng transgenic Cry1Ab protein/mL spiked whole blood.

Interlaboratory Evaluation of a New Reverse Transcriptase Polymerase Chain Reaction–Based Enzyme-Linked Immunosorbent Assay for the Detection of Circulating Melanoma Cells: A Multicenter Study of the Dermatologic Cooperative Oncology Group

2001 ◽  
Vol 19 (6) ◽  
pp. 1723-1727 ◽  
Author(s):  
U. Reinhold ◽  
C. Berkin ◽  
A.-K. Bosserhoff ◽  
A. Deutschmann ◽  
C. Garbe ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Tumor Cells ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Blood Samples ◽  
Melanoma Patients ◽  
Polymerase Chain ◽  
Interlaboratory Reproducibility

PURPOSE: Reverse transcription-polymerase chain reaction (RT-PCR)–based detection of tyrosinase mRNA is the most frequently used laboratory method for the detection of circulating tumor cells in melanoma patients. However, previously published results showed considerable variability in the PCR positivity rates. MATERIALS AND METHODS: We designed a collaborative study to assess the sensitivity, specificity, and clinical relevance of a new standardized RT-PCR–based enzyme-linked immunosorbent assay (ELISA) for the detection of circulating melanoma cells. Blood samples of healthy donors mixed with cells of a melanoma cell line were prepared in a blinded fashion, and aliquots were sent to seven participating laboratories experienced in RT-PCR. RESULTS: The results demonstrate a high sensitivity (1 melanoma cell/mL blood) and specificity (no false-negatives and 7.4% [2 of 28] false-positives) of the assay and a satisfactory rate of interlaboratory reproducibility. The analysis of aliquots of blinded samples derived from 60 melanoma patients identified tyrosinase mRNA in 17 of 60 (28.3%): three (20%) of 15 stage I patients, two (13.3%) of 15 stage II patients, five (35.7%) of 14 stage III patients, and seven (43.8%) of 16 stage IV patients. The interlaboratory reproducibility of positive samples, however, was extremely low and indicates the presence of low amounts of target mRNA. CONCLUSION: Reverse transcriptase-PCR ELISA has a high sensitivity and specificity for the detection of tyrosinase mRNA in peripheral blood cells. The low interlaboratory reproducibility for the detection of tumor cells in blood samples of melanoma patients, however, raises the question of relevance of this assay for clinical use.

scholarly journals Comparison of Blood Polymerase Chain Reaction and Enzyme-Linked Immunosorbent Assay for Detection of Mycobacterium Avium Subsp. Paratuberculosis Infection in Cattle and Sheep

2005 ◽  
Vol 17 (4) ◽  
pp. 354-359 ◽  
Author(s):  
Ramon A. Juste ◽  
Joseba M. Garrido ◽  
Marivi Geijo ◽  
Natalia Elguezabal ◽  
Gorka Aduriz ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Immunosorbent Assay ◽  
Ileocecal Valve ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fisher Exact Test ◽  
Chain Reaction ◽  
Blood Samples ◽  
Exact Test ◽  
Polymerase Chain ◽  
Cattle And Sheep

A study was carried out to compare the performance of enzyme-linked immunosorbent assay (ELISA) and blood polymerase chain reaction (PCR) for diagnosis of paratuberculosis in cattle and sheep. For cattle, a set of 278 samples from 1 paratuberculosis-affected Friesian farm was used; it included 80 ELISA-positive samples and 198 ELISA-negative samples from an age-matched group. Ninety-four samples were from heifers and 184 were from 2–5-year-old cows. The overall analysis showed a clear association (Fisher exact test [FET] P = 0.0049) but a weak negative agreement (45.3%, kappa = −0.1665 ± 0.0994) between the 2 tests. It reflected a moderate agreement among heifers (87.7%, kappa = 0.4471 ± 0.2435) and a moderate disagreement among cows (62.7%, kappa = −0.3670 ± 0.1057). For sheep, 496 blood samples from 53 Latxa dairy flocks were used; 180 of the blood samples were from dam/offspring pairs. The overall association between the 2 tests on ovine samples was strong (FET, P = 0.0005), whereas the agreement was low (kappa = 0.1622 ± 0.1188). There was slightly better agreement for ewes (kappa = 0.2135 ± 0.1992) than for lambs (kappa = 0.1193 ± 0.1301). There was also a highly unlikely proportion of dam/offspring positive results (FET, P < 0.0001, kappa = 0.6269 ± 0.1854). Four of 6 lambs that were necropsied 1 year after testing had paratuberculosis microscopic lesions in the ileocecal valve (3 lambs) or a PCR-positive result (4 lambs). These results suggest that blood PCR testing might be a potentially useful new approach in paratuberculosis diagnosis, especially in young animals.

scholarly journals Evaluation of Sandwich Enzyme-Linked Immunosorbent Assay and Reverse Transcription Polymerase Chain Reaction for the Diagnosis of Foot-and-Mouth Disease

Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Genome ◽  
Foot And Mouth Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Kappa Value ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Level Of Agreement

<b><i>Background:</i></b> Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. <b><i>Methods:</i></b> A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. <b><i>Results:</i></b> S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (<i>p</i> = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. <b><i>Conclusions:</i></b> The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.

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