Clinical Relevance of Annual Screening Using a Commercial Enzyme-Linked Immunosorbent Assay (SNAP 3Dx) for Canine Ehrlichiosis

2009 ◽  
Vol 45 (3) ◽  
pp. 118-124 ◽  
Author(s):  
Barbara C. Hegarty ◽  
Pedro Paulo Vissotto de Paiva Diniz ◽  
Julie M. Bradley ◽  
Leif Lorentzen ◽  
Edward Breitschwerdt
Keyword(s):  
Clinical Relevance ◽  
Deoxyribonucleic Acid ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Active Infection ◽  
Chain Reaction ◽  
Canine Ehrlichiosis ◽  
Annual Screening ◽  
Polymerase Chain ◽  
Positive Results

Eighty-six dogs were selected based upon Ehrlichia (E.) canis SNAP 3Dx positive results to determine clinical relevance of annual E. canis screening. Immunofluorescence assay showed 72 (84%) of 86 dogs were seroreactive for E. canis. Polymerase chain reaction (PCR) revealed that 12 (14%) of 86 dogs had Ehrlichia deoxyribonucleic acid; seven had E. canis, four had E. ewingii, and one was coinfected with E. chaffeensis and E. ewingii. Thrombocytopenia (<164,000 platelets/μL) was found in 28 (39%) of 72 dogs. In this study, thrombocytopenia was frequently detected in healthy Ehrlichia SNAP 3Dx-positive dogs, whereas active infection was infrequently confirmed by PCR. Therefore, treatment based upon screening results alone is not recommended.

Molecular Biology for the Clinician: Understanding Current Methods

2006 ◽  
Vol 42 (5) ◽  
pp. 326-335 ◽  
Author(s):  
Heather L. Chandler ◽  
Carmen M.H. Colitz
Keyword(s):  
Molecular Biology ◽  
Blot Analysis ◽  
Deoxyribonucleic Acid ◽  
Northern Blot Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Biochemical Assays ◽  
Basic Understanding ◽  
Polymerase Chain ◽  
Pcr Microarray

The basics of molecular biology involve the replication of deoxyribonucleic acid and its transcription and translation into proteins. Biochemical assays such as the Southern blot analysis, polymerase chain reaction (PCR), Northern blot analysis, reverse-transcriptase PCR, microarray technology, Western blot analysis, immunohistochemistry, enzyme-linked immunosorbent assay, and flow cytometry utilize various aspects of molecular biology. To understand these assays requires some basic understanding of the principles of molecular biology. This paper provides basic information on the methodology and techniques used in these assays.

scholarly journals Genotyping ofSalmonellaspp. on the basis of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)

2018 ◽  
Author(s):  
Calvin Jiksing ◽  
Normah Yusop ◽  
Farhan Nazaie Nasib ◽  
Kenneth Francis Rodrigues
Keyword(s):  
Polymerase Chain Reaction ◽  
Mycobacterium Tuberculosis ◽  
Immunosorbent Assay ◽  
Deoxyribonucleic Acid ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Pcr Polymerase Chain Reaction ◽  
Crispr Array ◽  
Polymerase Chain ◽  
Veterinary Laboratory

ABSTRACTAimsBacterial genotyping on the basis of the CRISPR array has been established inMycobacterium tuberculosiswith a method called spacer oligonucleotide typing (spoligotyping). The spoligotyping method had been widely used for both detection and typing ofM. tuberculosiscomplex bacteria. This present study aimed at determining if the CRISPR array inSalmonellaspp. could be applied to establish a correlationship between serogroup and the fingerprint generated by CRISPR typing.Methodology and resultsA total of 30 samples were obtained from Diagnostic Veterinary Laboratory, Kota Kinabalu, Sabah. Serogroup was determined on the basis of ELISA (enzyme-linked immunosorbent assay). Four different serogroups were identified which were serogroup B, C, D, and E. DNA (deoxyribonucleic acid) was extracted and PCR (polymerase chain reaction) was performed using primers which were designed to amplify the CRISPR array inSalmonellagenome. Our results indicate that there is a correlationship between serogroup obtained using ELISA and the profile generated by CRISPR typing.Conclusion, significance and impact of studyCRISPR typing has the potential to be applied for the genotyping ofSalmonella.

scholarly journals Occurrence of Leishmania (Leishmania) chagasi in a domestic cat (Felis catus) in Andradina, São Paulo, Brazil: case report

2010 ◽  
Vol 19 (4) ◽  
pp. 256-258 ◽  
Author(s):  
Willian Marinho Dourado Coelho ◽  
Valéria Marçal Felix de Lima ◽  
Alessandro Francisco Talamini do Amarante ◽  
Helio Langoni ◽  
Virgínia Bodelão Richini Pereira ◽  
...  
Keyword(s):  
Natural Infection ◽  
Immunofluorescence Assay ◽  
Felis Catus ◽  
Domestic Cat ◽  
Enzyme Linked Immunosorbent Assay ◽  
Leishmania Chagasi ◽  
Chain Reaction ◽  
Human Visceral Leishmaniasis ◽  
Polymerase Chain ◽  
The City

This work describes natural infection by Leishmania in a domestic cat where amastigote forms of the parasite were observed in the popliteal lymph node imprint. Positive and negative serological reactions were observed by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFA), respectively. Polymerase chain reaction (PCR) revealed that the nucleotide sequence of the sample was identical to Leishmania (L.) chagasi. This is the first report of the disease in felines of the city of Andradina, SP, an area considered endemic for canine and human visceral leishmaniasis.

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.

scholarly journals Occurrence of abortions induced by Neospora caninum in dairy cattle from Santa Catarina, southern Brazil

2017 ◽  
Vol 26 (3) ◽  
pp. 292-298 ◽  
Author(s):  
Cesar Augusto Barbosa de Macedo ◽  
Madlaine Frigo Silveira Barbosa de Macedo ◽  
Ana Carolina Miura ◽  
Alessandra Taroda ◽  
Sergio Tosi Cardim ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Dairy Cows ◽  
High Frequency ◽  
Neospora Caninum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Southern Brazil ◽  
Chain Reaction ◽  
Santa Catarina ◽  
Tissue Samples ◽  
Polymerase Chain

Abstract The aim of the present study was to investigate the occurrence of N. caninum associated with abortions of dairy cattle from Santa Catarina state, southern Brazil by using enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), and polymerase chain reaction (PCR). Blood from dairy cows that aborted along with intrathoracic fluid and tissue samples (brain, heart, liver, and lung) from their fetuses were collected and used for serology; PCR, histopathological, and immunohistochemistry (IHC) evaluations were also conducted. Twenty-one cows (51.2%) out of 41, and eight fetuses (26.7%) out of 30 were ELISA (HerdCheck, IDEXX) positive for N. caninum. Dams > 36 months of age had a higher risk of being serum positive than younger animals. PCR and IHC revealed that 38.8% (14/36) and 25.0% (9/36) of the fetuses were positive for N. caninum, respectively for each of the tests. Seropositive cows had a higher frequency of fetuses that were also positive by either intrathoracic fluid, PCR, or IHC. In summary, the present study observed a high frequency of N. caninum in abortions from dairy cows from southern Brazil, with a higher N. caninum prevalence found in cows that were older than 36 months. In addition, serology, PCR, and IHC should be used all together for better diagnosis of neosporosis in cattle.

Identification of Bacteroides forsythus in Subgingival Dental Plaque with the Aid of a Rapid PCR Method

1997 ◽  
Vol 76 (7) ◽  
pp. 1376-1380 ◽  
Author(s):  
J.H. Meurman ◽  
J. Wahlfors ◽  
A. Korhonen ◽  
P. Alakuijala ◽  
P. Väisänen ◽  
...  
Keyword(s):  
Dental Plaque ◽  
Subgingival Plaque ◽  
Chain Reaction ◽  
Rapid Pcr ◽  
Microbiological Methods ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Pre Treatment ◽  
Culture Positive ◽  
Positive Results

Bacteroides forsythus has been shown to be prevalent among patients with periodontitis. Conventional microbiological methods used to identify this bacterium, however, are laborious and time-consuming and are therefore not well-suited for screening purposes. We have developed a polymerase chain-reaction (PCR) method which is rapid, specific, and simple to perform and does not require other sample pre-treatment except a brief centrifugation. This method was applied to the detection of B. forsythus in subgingival plaque of 58 periodontitis patients. When compared with the results of conventional culturing, the PCR method always confirmed the culture-positive results, while none of the PCR negative samples was shown to be culture-positive. The PCR method appeared to give more than double the number of samples positive for B. forsythus than culturing (89.7% vs. 37.9%). The analysis requires less than 4 hrs to perform, and is specific only to B. forsythus and sensitive enough to detect fewer than 5 bacteria.

scholarly journals Peripheral Blood Leukocytes and Serum Nested Polymerase Chain Reaction Are Complementary Methods for Monitoring Active Cytomegalovirus Infection in Transplant Patients

2013 ◽  
Vol 24 (3) ◽  
pp. e69-e74 ◽  
Author(s):  
PD Andrade ◽  
MT Fioravanti ◽  
EBV Anjos ◽  
C De Oliveira ◽  
DM Albuquerque ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Positive Predictive Value ◽  
Predictive Value ◽  
Cytomegalovirus Infection ◽  
Clinical Symptoms ◽  
Peripheral Blood Leukocytes ◽  
Active Infection ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Polymerase Chain

BACKGROUND: Human cytomegalovirus is an important cause of morbidity and mortality in immunocompromised patients. Qualitative polymerase chain reaction (PCR) has proven to be a sensitive and effective technique in defining active cytomegalovirus infection, in addition to having low cost and being a useful test for situations in which there is no need for quantification. Real-time PCR has the advantage of quantification; however, the high cost of this methodology makes it impractical for routine use.OBJECTIVE: To apply a nested PCR assay to serum (sPCR) and to evaluate its efficiency to diagnose active cytomegalovirus infection compared with PCR of peripheral blood leukocytes (L-PCR).METHODS: Samples of 37 patients were prospectively evaluated. An internal control was created and applied to sPCR to exclude false-negative results.RESULTS: In total, 21 patients (57%) developed active cytomegalovirus infection. After analyzing the two methods for the diagnosis of active infection, higher sensitivity and negative predictive value of the L-PCR versus sPCR (100% versus 62%), and higher specificity and positive predictive value of sPCR versus L-PCR (81% versus 50% and 72%, respectively) were observed. Discordant results were observed in 11 patients who were L-PCR-positive but sPCR-negative for active cytomegalovirus infection, five of whom developed clinical symptoms of cytomegalovirus. Clinical symptoms were observed in 14 patients, 12 of whom were diagnosed with active infection by nested L-PCR (P=0.007) and seven by nested sPCR (P=0.02). Higher specificity and a positive predictive value for sPCR were observed.CONCLUSION: Nested L-PCR and sPCR were considered to be complementary methods for the diagnosis and management of symptomatic cytomegalovirus infection.

Studies on Diagnosis of Bovine Brucellosis by Immunocyto-chemistry and Immunohistochemistry

2021 ◽  
Author(s):  
K.S. Lakshmikanth ◽  
N.S. Sharma ◽  
D. Pathak ◽  
Paviter Kaur
Keyword(s):  
Biochemical Analysis ◽  
Placental Tissue ◽  
Diagnostic Techniques ◽  
Clinical Samples ◽  
Bovine Brucellosis ◽  
Reproductive Disorders ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Polymerase Chain ◽  
Positive Results

Background: Brucellosis is a major threat to livestock economy and an important zoonotic disease. A rapid and accurate diagnosis is a necessity to curb the spread and progress of the disease. The current study aimed to evaluate sensitivity of Immunocytochemistry and Immunohistochemistry methods for detection of Brucella spp.Methods: A total of 50 samples comprising of fetal stomach content, vaginal discharges and placenta were collected from cattle and buffaloes suffering from abortions and other reproductive disorders in and around Ludhiana, Punjab during the period 2017-2018. All the samples were processed for isolation and confirmed with biochemical analysis and Polymerase chain reaction (PCR). The isolates obtained and 43 clinical samples excluding placental samples were subjected to Immunocytochemistry (ICC). Immunohistochemistry (ICH) was performed on placental samples.Result: A total of four isolates were recovered from the screened samples. The four isolates also yielded positive results in Immunocytochemistry. Among the 43 clinical samples screened by Immunocytochemistry, five were positive, however only 3 isolates were recovered on isolation. A total of seven placental tissue samples were processed and subjected to immunohistochemistry. Of the three placental samples positive by immunohistochemistry, only one sample was isolated on culture. The results suggest that both immunocytochemistry and immunohistochemistry are sensitive diagnostic techniques in comparison to isolation.

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