qPCR detection of Mycobacterium leprae DNA in urine samples of leprosy patients using the Rlep gene target.

Background: Leprosy, a chronic infectious disease caused by Mycobacterium leprae, continues to pose a public health challenge in many parts of the world. Early and accurate diagnosis is crucial for effective treatment and prevention of disabilities associated with the disease. Molecular techniques such as PCR have demonstrated great potential as a diagnostic tool for directly detecting M. leprae DNA in different clinical samples, providing better sensitivity and specificity than conventional diagnostic techniques. The objective of this study was to measure the amount of M. leprae DNA in leprosy patients' urine samples using the Rlep gene target through qPCR. Methods: Different clinical samples such as smear, blood, and urine samples were collected from leprosy patients and healthy individuals. Leprosy patients were classified by the Ridley-Jopling classification. The Ziehl-Neelsen staining method was used for the slit skin smear (SSS) samples, and the bacteriological index (BI) was calculated for leprosy patients. DNA extraction and qPCR were performed for all three types of clinical samples using the Rlep gene target. Results: The Mycobacterial leprae DNA was successfully detected and quantified in all clinical samples across all types of leprosy among all the study groups using the Rlep gene (129 bp) target. The Rlep gene target was able to detect the presence of M. leprae DNA in 100% of urine, 96.1% of blood, and 92.2% of SSS samples of leprosy patients. Urine samples showed significant differences (p < 0.001) between the control and the different clinical forms and between borderline tuberculoid (BT) and pure neuritic leprosy (PNL) cases. There are significant differences in cycle threshold (Ct) values between control cases and clinical categories (p < 0.001), as well as specific differences within clinical categories (p < 0.001), reflecting the variability in bacterial load and detection sensitivity across different sample types and clinical manifestations of leprosy. Conclusion: Overall, this study's findings suggest that the qPCR technique can be used to detect M. leprae DNA in urine samples of leprosy patients using the Rlep gene target. It can also be used for diagnosing the disease and monitoring the effectiveness of anti-leprosy drugs, including multi-drug therapy (MDT), across various leprosy disease groups.

In silico approach to design a multi-epitopic vaccine candidate targeting the non-mutational immunogenic regions in envelope protein and surface glycoprotein of SARS-CoV-2.

The novel corona virus (COVID-19) is a causative agent for severe acute respiratory syndrome (SARS-CoV-2) and responsible for the current human pandemic situation which has caused global social and economic commotion. The currently available vaccines use whole viruses whereas there is scope for peptide based vaccines. Thus, the global raise in statistics of this infection at an alarming rate evoked us to determine a novel and effective vaccine candidate against SARS-CoV-2. To find the potential vaccine candidate targets, immunoinformatics approaches were used to analyze the mutations in the envelope protein and surface glycoprotein and determine the conserved region; further specific T-cell epitopes VSLVKPSFY, SLVKPSFYV, RVKNLNSSR, SEETGTLIV, LVKPSFYVY, LTDEMIAQY, YLQPRTFLL, RLFRKSNLK, SPRRARSVA, AEIRASANL, TLLALHRSY, YSRVKNLNS and FELLHAPAT and B-cells epitopes TLAILTALRLCAYCCN and AGTITSGWTFGAGAAL were identified. The 3 D structure of epitope was predicted, refined and validated. The molecular docking analysis of multi-epitope vaccine candidates with TLR receptors, predicted effective binding. Overall, using bioinformatics approach this multi-epitopic target facilitates the proof of concept for SARS-CoV-2 conserved epitopic vaccine design.Communicated by Ramaswamy H. Sarma.

Retrieval of foreign body from a postoperative defect in the mandible during the follow-up period: A bizarre occurrence.

Though reported cases of foreign bodies left intraoperatively in the oral cavity are very few, there is no case mentioned in the literature where foreign body was left behind during follow-up visits. Here, we present an operated case of unicystic ameloblastoma of mandibular ramus region, in which a needle hub was left at the operated site (cavity created because of wound dehiscence) during some of the follow-up visits, which was detected accidently by radiograph and later on retrieved. The case reported was because of negligence of trainee surgeons, might be because of overburden or because of minimal interest in these repeated follow ups. But, a trainee should understand that their work also has similar importance as that of surgeon's work.