75 COMPARISON OF THE DG29TM ENZYME-LINKED IMMUNOSORBENT ASSAY KIT COMPARED WITH TRANSRECTAL ULTRASONOGRAPHY FOR EARLY PREGNANCY DIAGNOSIS FOLLOWING TRANSFER OF JAPANESE BLACK CATTLE EMBRYOS

2012 ◽  
Vol 24 (1) ◽  
pp. 150 ◽  
Author(s):  
Y. Nakamura ◽  
M. Urakawa ◽  
A. Ideta ◽  
A. Shirasawa ◽  
Y. Oono ◽  
...  
Keyword(s):  
Negative Predictive Value ◽  
Clinical Application ◽  
Immunosorbent Assay ◽  
Early Pregnancy ◽  
Predictive Value ◽  
Transrectal Ultrasonography ◽  
Enzyme Linked Immunosorbent Assay ◽  
Japanese Black Cattle ◽  
Elisa Kit ◽  
Pregnancy Diagnosis

In commercial embryo transfer industry, accurate early detection of pregnant and nonpregnant cows plays a key role in achieving an optimum calving-to-conception interval. The DG29TM enzyme-linked immunosorbent assay (ELISA) kit (Conception, Animal Reproduction Technologies, Beaumont, Canada) measures the level of pregnancy-related glycoproteins in blood that are linked to pregnancy. Here, we compared the DG29 kit with transrectal ultrasonography (TU) to evaluate the possibility of the clinical application of the ELISA kit for early pregnancy diagnosis. Embryos recovered from superovulated Japanese Black cows were transferred to 110 recipient Holstein heifers on Day 6 to 8 of the oestrous cycle (oestrus = Day 0). Pregnancy was diagnosed between Day 29 and Day 40 by TU with a 5.0/7.5-MHz linear array transducer (Tringa, Pie Medical Equipment B.V., Maastricht, the Netherlands). Blood samples were collected from the tail vein or artery into vacuum serum tubes after TU and serum was separated and stored at –30°C until the ELISA was performed. The ELISA results were interpreted as positive (pregnant, >1000 pg mL–1) or negative (nonpregnant, <300 pg mL–1). Ninety-nine of the 110 heifers were judged as pregnant or nonpregnant by TU. Seventy-six of the 99 heifers were judged as pregnant, in which fetuses were visualised clearly in the uterine horn. The following measures, sensitivity, specificity, predictive value and accuracy of pregnancy outcomes based on the ELISA results, were assessed by comparing with those based on the definite TU results. The values for sensitivity and specificity were 100% (76/76) and 91.3% (21/23), the positive and negative predictive value were 97.4% (76/78) and 100% (21/21), respectively and accuracy was 98.0% (97/99). On the other hand, chorioallantoic fluids in 11 of the 110 heifers were detected by TU around Day 30 of gestation, but fetuses were not identified or were unclearly visualised, which suggests that the embryos died during the peri-implantation period after transfer. Ten of the 11 heifers were classified as pregnant by the ELISA, but only 3 heifers were identified as pregnant with reexamination by later TU, which indicates that the pregnancy-related glycoproteins residue from embryo mortality was detected by the DG29 kit. However, the negative predictive value of the DG29 kit was 100% in this study. In conclusion, except for early embryonic death, the DG29 kit was highly accurate and suitable for clinical application in early pregnancy determination following transfer of Japanese Black cattle embryos.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Ringworm Infection in Cattle

2013 ◽  
Vol 20 (8) ◽  
pp. 1150-1154 ◽  
Author(s):  
Elena Tatiana Băguţ ◽  
Ludivine Cambier ◽  
Marie-Pierre Heinen ◽  
Vasile Cozma ◽  
Michel Monod ◽  
...  
Keyword(s):  
Positive Predictive Value ◽  
Negative Predictive Value ◽  
Immunosorbent Assay ◽  
Predictive Value ◽  
Skin Lesions ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Predictive Values ◽  
Content Type ◽  
Ringworm Infection

ABSTRACTThe aim of this study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of ringworm infection in cattle. We used available recombinant forms ofTrichophyton rubrumdipeptidyl peptidase V (TruDppV) andT. rubrumleucin aminopeptidase 2 (TruLap2), which are 98% identical toTrichophyton verrucosumorthologues. Field serum samples from 135 cattle with ringworm infection, as confirmed by direct microscopy, fluorescence microscopy, and PCR, and from 55 cattle without any apparent skin lesions or history of ringworm infection that served as negative controls were used. Sensitivities, specificities, and positive and negative predictive values were determined to evaluate the diagnostic value of our ELISA. Overall, the ELISAs based on recombinant TruDppV and TruLap2 discriminated well between infected animals and healthy controls. Highly significant differences (P< 0.0001, Mann-Whitney U test) were noted between optical density values obtained when sera from infected versus control cattle were tested. The ELISA developed for the detection of specific antibodies against DppV gave 89.6% sensitivity, 92.7% specificity, a 96.8% positive predictive value, and a 78.4% negative predictive value. The recombinant TruLap2-based ELISA displayed 88.1% sensitivity, 90.9% specificity, a 95.9% positive predictive value, and a 75.7% negative predictive value. To the best of our knowledge, this is the first ELISA based on recombinant antigens for assessing immune responses to ringworm infection in cattle; it is particularly suitable for epidemiological studies and also for the evaluation of vaccines and/or vaccination procedures.

Tissue plasminogen activator as a novel diagnostic aid in acute pulmonary embolism

VASA ◽  
2014 ◽  
Vol 43 (6) ◽  
pp. 450-458 ◽  
Author(s):  
Julio Flores ◽  
Ángel García-Avello ◽  
Esther Alonso ◽  
Antonio Ruíz ◽  
Olga Navarrete ◽  
...  
Keyword(s):  
Pulmonary Embolism ◽  
Negative Predictive Value ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Predictive Value ◽  
Acute Pulmonary Embolism ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Utility ◽  
Tissue Plasminogen ◽  
D Dimer

Background: We evaluated the diagnostic efficacy of tissue plasminogen activator (tPA), using an enzyme-linked immunosorbent assay (ELISA) and compared it with an ELISA D-dimer (VIDAS D-dimer) in acute pulmonary embolism (PE). Patients and methods: We studied 127 consecutive outpatients with clinically suspected PE. The diagnosis of PE was based on a clinical probability pretest for PE and a strict protocol of imaging studies. A plasma sample to measure the levels of tPA and D-dimer was obtained at enrollment. Diagnostic accuracy for tPA and D-dimer was determined by the area under the receiver operating characteristic (ROC) curve. Sensitivity, specificity, predictive values, and the diagnostic utility of tPA with a cutoff of 8.5 ng/mL and D-dimer with a cutoff of 500 ng/mL, were calculated for PE diagnosis. Results: PE was confirmed in 41 patients (32 %). Areas under ROC curves were 0.86 for D-dimer and 0.71 for tPA. The sensitivity/negative predictive value for D-dimer using a cutoff of 500 ng/mL, and tPA using a cutoff of 8.5 ng/mL, were 95 % (95 % CI, 88–100 %)/95 % (95 % CI, 88–100 %) and 95 % (95 % CI, 88–100 %)/94 %), respectively. The diagnostic utility to exclude PE was 28.3 % (95 % CI, 21–37 %) for D-dimer and 24.4 % (95 % CI, 17–33 %) for tPA. Conclusions: The tPA with a cutoff of 8.5 ng/mL has a high sensitivity and negative predictive value for exclusion of PE, similar to those observed for the VIDAS D-dimer with a cutoff of 500 ng/mL, although the diagnostic utility was slightly higher for the D-dimer.

Can Osteopontin Be Considered a Biomarker for Endometriosis?

2017 ◽  
Vol 68 (9) ◽  
pp. 2132-2134
Author(s):  
Daniela Roxana Albu (Matasariu) ◽  
Elena Mihalceanu ◽  
Alina Pangal ◽  
Carmen Vulpoi ◽  
Mircea Onofriescu ◽  
...  
Keyword(s):  
Serum Level ◽  
Immunosorbent Assay ◽  
Pelvic Pain ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Multifactorial Disease ◽  
Elisa Kit ◽  
Healthy Women ◽  
Progesterone Treatment ◽  
Large Dispersion

Endometriosis is a multifactorial disease that is manifested by infertility and pelvic pain. The purpose of the study was to evaluate the effect of progesterone treatment on the serum level of osteopontin, a multipotent cytokine, in patients with endometriosis. The study was prospective and we evaluated osteopontin levels that were measured in the serum of 40 patients with endometriosis and 12 healthy women using a standardized Enzyme-Linked Immunosorbent Assay (ELISA) kit. Osteopontin seric levels were lower in endometriosis patients and increased after progesterone treatment. Because of the large dispersion of data even in the control group, we find the association between osteopontin and endometriosis questionable.

scholarly journals Prevalence of feline coronavirus antibodies in cats in Bursa province, Turkey, by an enzyme-linked immunosorbent assay

2009 ◽  
Vol 11 (10) ◽  
pp. 881-884 ◽  
Author(s):  
Annamaria Pratelli ◽  
Kadir Yesilbag ◽  
Marcello Siniscalchi ◽  
Ebru Yalçm ◽  
Zeki Yilmaz
Keyword(s):  
Immunosorbent Assay ◽  
Predictive Value ◽  
Gold Standard ◽  
Population Based ◽  
Standard Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
High Association ◽  
Gold Standard Test ◽  
Feline Coronavirus

Feline sera from Bursa province (Turkey) were assayed for coronavirus antibody using an enzyme-linked immunosorbent assay (ELISA). The study was performed on 100 sera collected from cats belonging to catteries or community shelters and to households. The serum samples were initially tested with the virus neutralisation (VN) test and the results were then compared with the ELISA. The VN yielded 79 negative and 21 positive sera but the ELISA confirmed only 74 as negative. The ELISA-negative sera were also found to be free of feline coronoviruses-specific antibodies by Western blotting. Using the VN as the gold standard test, ELISA had a sensitivity of 100% and a specificity of 93.6%, with an overall agreement of 95%. The Kappa (κ) test indicated high association between the two tests (κ=0.86, 95% confidence interval (CI) 0.743–0.980). The positive predictive value (PPV) was 0.8, and the negative predictive value (NPV) was 0.93. The prevalence of FCoV II antibodies in the sampled population based on the gold standard was 62% (95% CI 0.44–0.77) among multi-cat environments, and 4% (95% CI 0.01–0.11) among single cat households.

314 EARLY PREGNANCY DIAGNOSIS USING A NOVEL TRANSRECTAL ULTRASONOGRAPHY PROTOCOL IN JAPANESE BLACK COWS

2009 ◽  
Vol 21 (1) ◽  
pp. 254
Author(s):  
A. Gaja ◽  
C. Kubota ◽  
T. Kojima
Keyword(s):  
Corpus Luteum ◽  
Early Pregnancy ◽  
Transrectal Ultrasonography ◽  
Blood Collection ◽  
Sectional Area ◽  
Cross Sectional ◽  
Plasma Progesterone ◽  
Area Change ◽  
Pregnancy Diagnosis ◽  
Peripheral Plasma

The present study aims to establish a novel practical protocol for early pregnancy diagnosis in cows by using transrectal ultrasonography. The protocol is based on measurements of corpus luteum (CL) cross-sectional area (CL c-s area) change performed at 2 separate days before the coming estrus after AI. Fourteen cows were inseminated artificially, and transrectal ultrasonographical observation of the ovaries and blood collection for measurement of peripheral plasma progesterone (P4) concentration were carried out daily from Days 12 to 23 (Day 0 = the day of onset of estrus). Thereafter, cows were routinely diagnosed for pregnancy at Day 30 by transrectal ultrasonography. The largest CL c-s area was obtained at Day 14 in both pregnant and non-pregnant cows. Seven out of 8 non-pregnant cows showed significant CL c-s area regression between Days 14 and 20 (422 ± 112 v. 249 ± 63 mm2), whereas no regression was observed between Days 14 and 20 in pregnant cows (416 ± 65 v. 402 ± 78 mm2). The regression in the CL c-s area between pregnant and non-pregnant cows was significantly different during Day 18 (424 ± 65 v. 288 ± 88 mm2) to Day 23 (402 ± 71 v. 139 ± 64 mm2). P4 concentration was significantly low (less than 1 ng mL–1) at Day 20 in 3 out of 8 non-pregnant cows, whereas the pregnant cows showed significant increase of P4 between Days 14 and 20 (2.6 ± 0.2 v. 3.4 ± 0.5 ng mL–1). The pregnant cows showed significantly higher P4 concentration starting from Day 18 than non-pregnant cows. However, in non-pregnant cows, 4 cows returned to estrus on Day 20 or after, 3 cows showed no signs of estrus, and 1 cow came in estrus as early as Day 18 after AI. In conclusion, the results of the present study suggest that measuring the change in the CL c-s area at Days 14 and 20 makes it possible to detect the non-pregnant cows at Day 20 after AI. However, it was also indicated that measuring the change of P4 concentrations on the same days did not always successfully detect non-pregnant cows. The new protocol based on CL c-s area regression rate can detect almost certainly non-pregnant cows at Day 20 after AI. It is suggested that this method is advantageous in research and industrial breeding.

scholarly journals Determination of Prolactin in Canine Saliva: Is it Possible to Use a Commercial ELISA kit?

Animals ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 418 ◽  
Author(s):  
Gutiérrez ◽  
Gazzano ◽  
Torracca ◽  
Meucci ◽  
Mariti
Keyword(s):  
Stress Response ◽  
Immunosorbent Assay ◽  
Matrix Effect ◽  
Enzyme Linked Immunosorbent Assay ◽  
Blood Samples ◽  
Elisa Kit ◽  
Cases Study ◽  
Dog Blood

Prolactin has been reported to be a remarkable index of stress response, both acute and chronic, in several species. The use of biological matrixes other than blood is receiving increasing interest in the study of hormones, due to the lower invasiveness in collection. This research aimed to investigate the possibility of using a commercial ELISA (enzyme-linked immunosorbent assay) kit for measuring canine prolactin in blood for the quantification of canine prolactin in saliva. Study 1 consisted of a validation protocol, using saliva samples collected from lactating and non-lactating dogs. Study 2 was conducted to investigate a possible correlation between prolactin concentration in saliva and plasma in sheltered dogs by using the same kit. Prolactin values were reliably read only when they came from blood samples, not from saliva, but tended to be low in most of the cases. Study 1 showed that saliva had a matrix effect. In study 2, saliva prolactin levels were low and in 42.9% of cases, not readable. No correlation between prolactin values in plasma and saliva was found (ρ=0.482; p=0.274). These findings suggested that the determination of prolactin in dog saliva through an ELISA kit created for measuring prolactin in dog blood was unreliable.

scholarly journals Clinical chorioamnionitis at term VI: acute chorioamnionitis and funisitis according to the presence or absence of microorganisms and inflammation in the amniotic cavity

2015 ◽  
Vol 0 (0) ◽  
Author(s):  
Roberto Romero ◽  
Piya Chaemsaithong ◽  
Nikolina Docheva ◽  
Steven J. Korzeniewski ◽  
Juan P. Kusanovic ◽  
...  
Keyword(s):  
Negative Predictive Value ◽  
Predictive Value ◽  
Acute Inflammation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Likelihood Ratios ◽  
Pediatric Pathology ◽  
Amniotic Cavity ◽  
Fluid Analysis ◽  
Increased Risk ◽  
Histologic Chorioamnionitis

AbstractNeonates born to mothers with clinical chorioamnionitis at term are at an increased risk of infection. Acute subchorionitis, chorioamnionitis, and funisitis are considered placental histologic features consistent with acute inflammation according to the Society for Pediatric Pathology. The objectives of this study were to examine the performance of placental histologic features in the identification of: 1) microbial-associated intra-amniotic inflammation (intra-amniotic infection); and 2) fetal inflammatory response syndrome (FIRS).This retrospective cohort study included women with the diagnosis of clinical chorioamnionitis at term (n=45), who underwent an amniocentesis to determine: 1) the presence of microorganisms using both cultivation and molecular biologic techniques [polymerase chain reaction (PCR) with broad range primers]; and 2) interleukin (IL)-6 concentrations by enzyme-linked immunosorbent assay (ELISA). The diagnostic performance (sensitivity, specificity, accuracy, and likelihood ratios) of placental histologic features consistent with acute inflammation was determined for the identification of microbial-associated intra-amniotic inflammation and FIRS.1) The presence of acute histologic chorioamnionitis and funisitis was associated with the presence of proven intra-amniotic infection assessed by amniotic fluid analysis; 2) funisitis was also associated with the presence of FIRS; 3) the negative predictive value of acute funisitis ≥stage 2 for the identification of neonates born to mothers with intra-amniotic infection was <50%, and therefore, suboptimal to exclude fetal exposure to bacteria in the amniotic cavity; and 4) acute funisitis ≥stage 2 had a negative predictive value of 86.8% for the identification of FIRS in a population with a prevalence of 20%.Acute histologic chorioamnionitis and funisitis are associated with intra-amniotic infection and the presence of FIRS. However, current pathologic methods have limitations in the identification of the fetus exposed to microorganisms present in the amniotic cavity. Further studies are thus required to determine whether molecular markers can enhance the performance of placental pathology in the identification of neonates at risk for neonatal sepsis.

A study of the correlation between Coxiella burnetii seropositivity and abortions in sheep in Eastern and Southeastern Turkey

2016 ◽  
Author(s):  
Ayse Kilic ◽  
Hakan Kalender
Keyword(s):  
Immunosorbent Assay ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Eastern Anatolia ◽  
Obligate Intracellular Bacterium ◽  
Serum Samples ◽  
Obligate Intracellular ◽  
Elisa Kit

Q fever is a zoonotic disease that occurs worldwide and is caused by the obligate intracellular bacterium Coxiella burnetii. Infected animals are usually asymptomatic, but infection can cause abortion and stillbirth in ruminants. The main purpose of this study was to evaluate prevalance of Coxiella burnetii infection in aborted and nonaborted sheep serum samples in Eastern Anatolia region by using enzyme-linked immunosorbent assay (ELISA). The determine of prevalance in sheep flocks from four provinces (Elazig, Malatya, Tunceli, Bitlis) and tested for anti-C.burnetii antibody detection, by means of Chekit Q fever Elisa kit. 350 serum samples obtained from flocks belonging aborted sheep showed that a total of 56 (16%) were detected seropositivity, whereas 171 serum samples obtained from nonaborted sheep flocks in 13 of the 171 (7.60%) for C.burnetii in seropositivity were observed. Coxiellosis should be considered an important cause of sheep with abortion history and nonaborted in Elazig and neighboring provinces.

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