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Introduction: Herpes Simplex Virus-1 (HSV-1) and HSV-2 can produce mucocutaneous lesions and Central Nervous System (CNS) infections, some of which may be life-threatening. There are no known animal vectors for HSV, and although experimental animals are easily infected, humans appear to be the only natural reservoir. Herpes infection is the predominant cause of genital ulcers worldwide. An increase in Polymerase Chain Reaction (PCR) usage for HSV detection and an increase in HSV-2 reactivation frequency among HSV/Human Immunodeficiency Virus (HIV)-co-infected persons can be attributed to this. Aim: To detect HSV infection by light microscopy, Enzyme Linked Immuno Sorbent Assay (ELISA), PCR in clinically suspected cases and to assess the utility value of PCR over ELISA and light microscopy. Materials and Methods: A cross-sectional study was conducted on 84 samples collected from clinically suspected cases of herpes simplex infections at the Department of Dermatology and Sexually Transmitted Diseases (STD) Clinic. The collected samples were processed in the Department of Microbiology, Bangalore Medical College and Research Institute, Bengaluru, Karnataka, India during the period from November 2018 to May 2020. Swabs collected from the patients were processed for investigations including light microscopy, HSV-1, HSV&2 PCR. Blood samples were collected from those patients for HSV1+2 IgM ELISA. Descriptive statistics such as mean and Standard Deviation (SD) for continuous variables, and frequencies and percentages for categorical variables were calculated. Results: Total of 84 cases of HSV were included 68 (81%) were with suspected cases of herpes genitalis, and 16 (19%) presented with herpes labialis. The prevalence of HSV infection was found to be 59.5%. In the case of herpes genitalis and herpes labialis, the majority were in the age group 31-40 years. Males were more affected compared to females in the case of herpes genitalis, while females were more affected in the case of herpes labialis. PCR had a sensitivity, specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV) of 74%, 100%, 100%, and 72.3%, respectively. Conclusion: PCR can be used as a gold standard test as well as a confirmatory test when compared to IgM ELISA and Tzanck smear for HSV diagnosis.

Introduction: Bacterial Sepsis by Multidrug Resistant Gram Negative Bacilli (MDRGNB) producing Extended Spectrum β-Lactamases (ESBL) is one of the major causes of mortality and morbidity in hospitals. Early detection of ESBLs directly from positive blood cultures can reduce mortality. The phenotypic detection of ESBLs is difficult as they may be masked by the co-production of additional enzymes like AmpC. This can be overcome by using an Aztreonam Discs With and Without Clavulanate (AO/CL) method. Aim: To identify ESBLs directly from the positive blood cultures by using AO/CL disc diffusion method and to detect the genes coding for ESBL enzymes by conventional Polymerase Chain Reaction (PCR). Materials and Methods: A prospective study was conducted over a period of five months (October 2020-February 2021). A total of 100 positive blood cultures showing Gram negative bacilli on Gram stain was subjected to direct detection of ESBLs by using Cefotaxime (CTX), Ceftazidime (CAZ) discs with and without clavulanate and AO/CL. Isolates from positive blood culture were identified to genus and species level by VITEK-2 compact. Isolates were tested for ESBL production by CAZ/ CTX with and without clavulanate disc diffusion method as recommended by CLSI. PCR was carried out to detect target genes responsible for ESBL production such as CTX–M, TEM, SHV genes. Statistical analysis was done by using MS Excel sheet. Descriptive statistics like percentage calculation was done in the study. Results: Out of 100 positive blood cultures showing Gram Negative Bacteria (GNB) on Gram stain, 33 were positive for ESBL production by direct disc diffusion method. Out of these, 27 ESBL producers were detected by CAZ/CTX with and without clavulanate disc diffusion method and AO/CL method whereas 6 ESBL producers were detected by AO/ CL disc diffusion method only. A 27 culture isolates were found positive for ESBL production by CAZ/CTX with and without clavulanate disc diffusion method as recommended by Clinical and Laboratory Standards Institute (CLSI). Out of 33, 28 (85%) isolates possessed one of the target genes for ESBL production such as 10TEM (36%), 10CTX-M (36%), 07TEM+CTX M (25%), 01SHV (3%). Conclusion: Direct detection of ESBLs plays a significant role in management of sepsis. It helps the clinician in escalation and de-escalation of antibiotics and prevents the development of antimicrobial resistance. It contributes towards antibiotic stewardship and better compliance to infection prevention and control protocols. AO/CL method is preferred to detect ESBL producers directly from positive blood culture bottles.