scholarly journals Sensitivity and specificity levels of two rapid assays for antibodies to Anaplasma spp. in dogs

2017 ◽  
Vol 30 (2) ◽  
pp. 290-293 ◽  
Author(s):  
Jiayou Liu ◽  
Matthew Eberts ◽  
Hannah Bewsey ◽  
Thomas P. O’Connor ◽  
Ramaswamy Chandrashekar ◽  
...  

Canine anaplasmosis is a tick-borne disease of dogs that results following infection with Anaplasma phagocytophilum or Anaplasma platys. The SNAP 4Dx Plus test (IDEXX Laboratories) and the VetScan Canine Anaplasma Rapid test (Abaxis) are commercial in-house rapid tests for the detection of antibody to these 2 antigenically related Anaplasma species. We evaluated 2 tests using serum and whole blood samples obtained from reference laboratories and veterinary hospitals. Samples were obtained from regions of the country known to be habitats of the primary tick vectors. The A. phagocytophilum sample set comprised 236 dog sera from the northeastern and midwestern United States; the A. platys sample set comprised 179 sera from dogs living in the southwestern United States. An indirect immunofluorescent antibody (IFA) test and an A. platys species-specific ELISA were used as reference assays for the A. phagocytophilum and A. platys samples, respectively. The SNAP test demonstrated significantly higher sensitivity (84.7% for A. phagocytophilum and 83.1% for A. platys), compared to the VetScan test (39.0% for A. phagocytophilum and 57.6% for A. platys). The specificity of the SNAP test (95.8% for A. phagocytophilum and 99.2% for A. platys) was significantly greater than the VetScan test (85.6% for A. phagocytophilum and 82.5% for A. platys). In a separate clinic study, conducted within an A. phagocytophilum–endemic state (Minnesota) using 154 whole blood samples from client-owned dogs, the VetScan test was negative for 22 of 39 SNAP and IFA seropositive samples.

2020 ◽  
Vol 38 (3) ◽  
pp. 222
Author(s):  
David Ardiyanto ◽  
Hastari Wuryastuty ◽  
Raden Wasito

Abstract              Brucellosis is a zoonotic disease that cause a significant economic losses for cattle industries worldwide. A rapid, precise and accurate diagnosis technique for diagnosis of brucellosis in all stages of the infection is definitely required.  Blood-samples are widely used for PCR-based DNA analysis because they are easily collected, handled, and processed. Direct PCR analysis without DNA extraction has been attempted to reduce time and  costs for routine analysis. This approach is promising but is still limited by the presence of PCR inhibitors that is naturally found  in the blood samples. The objective of this study was to compare the effectivity of direct PCR technique with or without DNA extraction for detection of Brucella abortus in the blood samples. Three whole-blood samples from brucella infected dairy cattle and five whole-blood samples  from beef cattle that having abortion were used as samples in this study. A pair of  bcsp31 primers and IS711 primers were used for amplification of genus-specific and species-specific of Brucella.  The results showed that amplicon in the position of 223 bp and 498 bp that are specific for B. abortus were detected from all of the samples that were analyzed on 1.5% agarose gels. Based on the result it could be concluded that direct PCR analyses without DNA extraction is a sensitive, specific, simple, rapid  and inexpensive assay for detecting B. abortus in the whole blood samples for either dairy or beef cattle and therefore it could  improve the existing surveillance and control programs for brucellosis. Keywords : brucellosis; direct PCR; PCR inhibitor; whole-blood sample; without DNA extraction                           Abstrak              Brucellosis adalah penyakit zoonosis yang menyebabkan kerugian ekonomi yang signifikan bagi industri ternak di seluruh dunia. Teknik diagnosis yang cepat, tepat dan akurat yang dapat digunakan untuk diagnosis brucellosis pada semua tahap infeksi sangat diperlukan. Sampel darah banyak digunakan untuk analisis PCR berbasis DNA karena mudah untuk dikoleksi, ditangani, dan diproses. Metoda PCR langsung tanpa didahului dengan ekstraksi DNA dikembangkan dengan tujuan penghematan waktu dan beaya untuk analisa secara rutin. Tehnik ini sangat menjanjikan tetapi memiliki keterbatasan karena adanya senyawa penghambat PCR yang secara alami terkandung di dalam sampel darah . Tujuan dari penelitian ini adalah membandingkan efektifitas antara uji PCR secara langsung dengan ekstraksi dan tanpa ekstraksi DNA untuk deteksi Brucella abortus di dalam darah. Tiga ( 3 ) sampel darah-EDTA yang berasal dari  sapi penderita brucellosis dan 5 sampel darah-EDTA dari sapi potong yang mengalami abortus digunakan sebagai sampel dalam penelitian ini. Pasangan primer bcsp31 dan primer IS711 untuk amplifikasi gen dan species specific digunakan dalam penelitian. Hasil menunjukkan bahwa amplikon/pita pada posisi 223 bp dan 498 bp yang spesifik untuk Brucella abortus terdeteksi dari semua sampel yang dianalisa dengan gel agarosa 1,5%. Berdasarkan hasil penelitian dapat disimpulkan bahwa uji PCR secara langsung tanpa didahului dengan ekstraksi DNA merupakan tehnik yang sensitif, spesifik, sederhana, cepat dan murah untuk deteksi B. abortus di dalam sampel darah baik sapi perah maupun sapi potong dan oleh karena itu diharapkan dapat digunakan untuk memperbaiki program kontrol dan survailance yang telah ada untuk brucellosis. Kata kunci : brucellosis; PCR langsung; penghambat PCR; sampel darah-utuh; tanpa ekstraksi DNA


2010 ◽  
Vol 41 (02) ◽  
Author(s):  
N Shazi ◽  
A Böss ◽  
HJ Merkel ◽  
F Scharbert ◽  
D Hannak ◽  
...  

Separations ◽  
2021 ◽  
Vol 8 (6) ◽  
pp. 78
Author(s):  
Sevasti Karampela ◽  
Jessica Smith ◽  
Irene Panderi

An ever-increasing need exists within the forensic laboratories to develop analytical processes for the qualitative and quantitative determination of a broad spectrum of new psychoactive substances. Phenylethylamine derivatives are among the major classes of psychoactive substances available on the global market and include both amphetamine analogues and synthetic cathinones. In this work, an ultra-high-performance liquid chromatography-positive ion electrospray ionization tandem mass spectrometric method (UHPLC-ESI-MS/MS) has been developed and fully validated for the determination of 19 psychoactive substances, including nine amphetamine-type stimulants and 10 synthetic cathinone derivatives, in premortem and postmortem whole blood. The assay was based on the use of 1 mL premortem or postmortem whole blood, following solid phase extraction prior to the analysis. The separation was achieved on a Poroshell 120 EC-C18 analytical column with a gradient mobile phase of 0.1% formic acid in acetonitrile and 0.1% formic acid in water in 9 min. The dynamic multiple reaction monitoring used in this work allowed for limit of detection (LOD) and lower limit of quantitation (LOQ) values of 0.5 and 2 ng mL−1, respectively, for all analytes both in premortem and postmortem whole blood samples. A quadratic calibration model was used for the 12 quantitative analytes over the concentration range of 20–2000 ng mL−1, and the method was shown to be precise and accurate both in premortem and postmortem whole blood. The method was applied to the analysis of real cases and proved to be a valuable tool in forensic and clinical toxicology.


2021 ◽  
Vol 2 (1) ◽  
pp. 100311
Author(s):  
Daniella C. Terenzi ◽  
Ehab Bakbak ◽  
Justin Z. Trac ◽  
Mohammad Al-Omran ◽  
Adrian Quan ◽  
...  

Author(s):  
Polina A. Dyachenko Timoshina ◽  
Leonid E. Dolotov ◽  
Ekaterina N. Lazareva ◽  
Anastasiia A. Kozlova ◽  
Olga A. Inozemtseva ◽  
...  

1994 ◽  
Vol 42 (3) ◽  
pp. 231-241 ◽  
Author(s):  
C. Shenberg ◽  
S. Spiegel ◽  
S. Chaitchik ◽  
P. Jordan ◽  
M. Kitzis ◽  
...  

1978 ◽  
Vol 24 (12) ◽  
pp. 2135-2138 ◽  
Author(s):  
K W Jackson

Abstract Each of 65 laboratories analyzed 10 whole-blood samples for erythrocyte protoporphyrin by one or more of several analytical procedures. These procedures were of two types: (a) extraction of protoporphyrin from the erythrocytes into ethyl acetate/acetic acid, re-extraction into hydrochloric acid, and fluorometric measurement; or (b) direct reading in a portable fluorometer (hematofluorometer), with no pretreatment of the blood sample. Interlaboratory correlation was generally poor, especially between laboratories using extraction procedures. Hematofluorometric results intercorrelated better, but they had a low bias as compared to the extraction approach. Nationwide standardization of the test is required to assure satisfactory interlaboratory performance and to identify laboratories whose results are sufficiently accurate to be used for interpretations according to guidelines set forth by the Center for Disease Control for erythrocyte protoporphyrin testing.


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