Evaluation of the diagnostic potential and DIVA capability of recombinant LigBCon1-5 protein of Leptospira interrogans serovar Pomona in canine leptospirosis.

BACKGROUND: Canine leptospirosis is a serious public health concern. AIMS: This study aims to investigate the feasibility of conserved first to fifth domains of recombinant Leptospira immunoglobulin like protein B antigen (rLigBCon1-5) as a serodiagnostic marker for detecting canine leptospirosis. METHODS: A total of 340 unvaccinated canine serum samples were screened using microscopic agglutination test (MAT) and rLigBCon1-5 based immunoglobulin G (IgG) indirect-enzyme-linked immunosorbent assay (I-ELISA). Further, 60 vaccinated canine sera were screened using MAT and rLigBCon1-5 based latex agglutination test (LAT). RESULTS: Microscopic agglutination test results revealed seropositivity of 28.6%. The relative sensitivity, specificity, and accuracy of IgG I-ELISA in comparison to MAT were 100%, 96.0%, and 97.2%, respectively. Out of 60 vaccinated sera, 46 sera reacted with MAT alone, and eight sera reacted by both tests, while six sera were non-reactive with both tests. Anti-LigB antibodies were detected in eight canine sera by rLigBCon1-5 based LAT. In five LAT reactive sera, agglutinins of locally circulating Leptospira serovars Grippotyphosa (n=4) and Australis (n=1) were detected. In three LAT reactive sera, agglutinins against Icterohaemorrhagiae (n=3) produced due to natural infection were present. CONCLUSION: Immunoglobulin G based indirect ELISA assay (IgG I-ELISA) can be employed as an alternative test instead of MAT. rLigBCon1-5 based LAT detected anti-LigB antibodies in eight vaccinated sera where the vaccine failure occurred partially or totally due to the limited efficacy spectrum of Nobivac® RL and cold chain breakage. This vaccine could not provide cross-protection against locally circulating Leptospira serovars. The recombinant LigBCon1-5 antigen based LAT possesses capability of differentiating infected from vaccinated individuals (DIVA capability) when employed as a pen-side test for detecting canine leptospirosis.

Genetic predisposition and prediction protocol for epithelial neoplasms in disease-free individuals: A systematic review.

BACKGROUND: Epithelial neoplasm is an important global health-care problem, with high morbidity and mortality rates. Early diagnosis and appropriate treatment are essential for increased life survival. Prediction of occurrence of malignancy in a disease-free individual by any means will be a great breakthrough for healthy living. AIMS AND OBJECTIVES: The aims and objectives were to predict the genetic predisposition and propose a prediction protocol for epithelial malignancy of various systems in our body, in a disease-free individual. METHODS: We have searched databases both manually and electronically, published in English language in Cochrane group, Google search, MEDLINE and PubMed from 2000 to 2019. We have included all the published, peer-reviewed, narrative reviews; randomized controlled trials; case-control studies; and cohort studies and excluded the abstract-only articles and duplicates. Specific words such as "etiological factors," "pathology and mutations," "signs and symptoms," "genetics and IHC marker," and "treatment outcome" were used for the search. A total of 1032 citations were taken, and only 141 citations met the inclusion criteria and were analyzed. RESULTS: After analyzing various articles, the etiological factors, clinical signs and symptoms, genes and the pathology involved and the commonly used blood and tissue markers were analyzed. A basic investigation strategy using immunohistochemistry markers was established. CONCLUSION: The set of proposed biomarkers should be studied in future to predict genetic predisposition in disease-free individuals.

Prospective evaluation of the diagnostic potential of LipL32 based latex agglutination test for Bovine leptospirosis.

The Livestock Sector of India plays an important role in livelihood security and socioeconomic development of rural households. Leptospirosis is an important zoonotic disease responsible for septicaemia, interstitial nephritis, jaundice, abortion, reproductive problem in most of the animal species. Reproductive disturbances in bovine population is most often restricted to investigation of brucellosis, however apart from brucellosis, there are many undiagnosed diseases like leptospirosis that takes a toll in the reproductive anomalies of cattle and buffalo. Hence, the present study was elucidated to screen the seroprevalence of Leptospira in cattle and buffalo in various hamlets of North India using a user friendly screening test i.e. LipL32 latex agglutination test. The overall seropositivity was found to be 26.01% (230/884) in case of bovine in this study and the LipL32 LAT showed a profound sensitivity and specificity with level of 94.97% and 99.53% respectively.

Prospective evaluation of the diagnostic potential of LipL32 based latex agglutination test for Bovine leptospirosis

@#The Livestock Sector of India plays an important role in livelihood security and socioeconomic development of rural households. Leptospirosis is an important zoonotic disease responsible for septicaemia, interstitial nephritis, jaundice, abortion, reproductive problem in most of the animal species. Reproductive disturbances in bovine population is most often restricted to investigation of brucellosis, however apart from brucellosis, there are many undiagnosed diseases like leptospirosis that takes a toll in the reproductive anomalies of cattle and buffalo. Hence, the present study was elucidated to screen the seroprevalence of Leptospira in cattle and buffalo in various hamlets of North India using a user friendly screening test i.e. LipL32 latex agglutination test. The overall seropositivity was found to be 26.01% (230/884) in case of bovine in this study and the LipL32 LAT showed a profound sensitivity and specificity with level of 94.97% and 99.53% respectively.

Rapid and visual detection of Leptospira in urine by LigB-LAMP assay with pre-addition of dye.

Leptospirosis is considered to be the most widespread zoonotic disease caused by pathogenic species of Leptospira. The present study reports a novel set of primers targeting LigB gene for visual detection of pathogenic Leptospira in urine samples through Loop-mediated isothermal amplification (LAMP). The results were recorded by using Hydroxyl napthol blue (HNB), SYBR GREEN I and calcein. Analytical sensitivity of LAMP was as few as 10 leptospiral organisms in spiked urine samples from cattle and dog. LigB gene based LAMP, termed as LigB-LAMP, was found 10 times more sensitive than conventional PCR. The diagnostic specificity of LAMP was 100% when compared to SYBR green qPCR for detection of Leptospira in urine samples. Though qPCR was found more sensitive, the rapidity and simplicity in setting LAMP test followed by visual detection of Leptospira infection in clinical samples makes LigB-LAMP an alternative and favourable diagnostic tool in resource poor setting.

Assessment of histological changes in antemortem gingival tissues fixed at various time intervals: A method of estimation of postmortem interval.

INTRODUCTION: Conventional methods to estimate the time of death are adequate, but a histological method is yet unavailable to assess postmortem interval (PMI). The autolytic changes that occur in an unfixed antemortem gingival tissue which reflects histologically at an early stage are similar to changes that occur in postmortem tissue. These histological changes can be used and applied in a postmortem tissue as a method to assess PMI. AIMS: The aim of the study is to assess the histological changes in a gingival tissue left unfixed for various time intervals and to correlate the findings with duration. MATERIALS AND METHODS: Sixty gingival tissues obtained from patients following therapeutic extractions, impactions, gingivectomy and crown lengthening procedures were used. Each tissue obtained was divided into two pieces and labeled as "A", the control group and " B" the study group. Tissues labeled "A" were fixed in 10% formalin immediately and tissues labeled"B" were placed in closed containers and fixed after 15, 30, 45 min, 1, 2, and 4 h time interval. Of the sixty tissues in the study group " B", ten tissues were used for each time interval under investigation. All the fixed tissues were processed, stained, assessed, and analyzed statistically using Pearson correlation and regression analysis. RESULTS: Histological changes appear at 15 min in an unfixed antemortem tissue. At 2 h interval, all layers with few cells in basal cell layer are involved. At 4 h interval, loss of stratification and complete homogenization of cells in the superficial layers with prominent changes in basal layer is evident. There was a positive correlation (<1.0) between the time interval and the appearance of the histological changes. CONCLUSION: Histological changes such as complete homogenization of cells in superficial layers and loss of epithelial architecture at 4 h in unfixed antemortem tissue may be used as a criterion to estimate PMI, after further studies on postmortem tissues.

Stevens-Johnson syndrome induced by a combination of lamotrigine and valproic acid.

Lamotrigine and valproic acid are well-tolerated anticonvulsants, but frequently associated with severe cutaneous reactions, such as the Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis, when used in combination. We report a case of SJS likely induced by the use of a lamotrigine and valproic acid regimen and as a dental surgeon it is important to identify such lesion and report to pharmacovigilance.

Evaluation of recombinant LigB antigen-based indirect ELISA and latex agglutination test for the serodiagnosis of bovine leptospirosis in India.

Leptospirosis is a zoonotic disease caused by pathogenic spirochetes of the genus Leptospira, causing febrile infection characterized by multi-organ failure in humans and animals. Leptospiral Ig-like protein B (LigB) is a surface-expressed antigen that mediates host cell invasion or attachment. In this study, N-terminal conserved region of LigB protein (46 kDa) was evaluated for its diagnostic potential to detect anti-leptospiral antibodies in the sera of various animal species. Dot blot analysis revealed immunoreactivity of Leptospira-positive sera of cattle, buffalo, dog, sheep and goat to purified LigB protein. We have analyzed 1126 bovine serum samples, collected from Northern and Eastern part of India, by microscopic agglutination test (MAT) and recombinant LigB (rLigB) based ELISA and latex agglutination test (LAT). The sensitivity of rLigB based ELISA for 554 MAT positive sera was 96.9% and the specificity with 572 MAT negative sera was 91.08% whereas LAT showed sensitivity and specificity of 93.68% and 92.31%, respectively. Kappa values of 0.879 and 0.860 for recombinant antigen based ELISA and LAT indicate excellent agreement with the gold standard serological test, MAT, for the detection of anti-leptospiral antibodies in sera. Further, LAT based on rLigB antigen is a simple and rapid test, suitable for serodiagnosis of leptospirosis under field conditions, owing to its portability and longer shelf life.