Leptospirosis in central & eastern Uttar Pradesh, an underreported disease: A prospective cross-sectional study.

Background & objective: Leptospirosis is a zoonotic disease associated with potentially fatal consequences and a grossly underreported disease in Uttar Pradesh. However, only a few studies are available which report the prevalence of leptospirosis in this State. Hence, this study was undertaken to know the status of the disease in central and eastern Uttar Pradesh. Methods: A total of 143 serum and urine samples were collected from patients with acute febrile illness from July 2017 to March 2019. All the serum samples were tested for Leptospira by rapid IgM antibody card and IgM ELISA and urine samples were tested by real-time polymerase chain reaction (RT-PCR) to detect Leptospira DNA. All positive and 10 per cent negative sera from ELISA and RT-PCR (all rapid test positive were also ELISA positive) were sent to the ICMR-Regional Medical Research Centre, Port Blair for microscopic agglutination test (MAT). Results: Thirty eight (26.6%) out of 143 samples were positive for leptospirosis either by ELISA or RT-PCR. Positive results were eight (6%) by Rapid card, 32 (22%) by IgM ELISA, 10 (7%) by MAT, 10 (7%) by RT-PCR. In MAT, the most common serovar was Lai followed by Hebdomadis, Bangkinang and Pomona. Interpretation & conclusions: Leptospirosis was found to be one of the important causes for acute febrile illness in the central and eastern parts of Uttar Pradesh. The results of the present study suggest that it is necessary to increase diagnostic facility and awareness in clinicians for the screening of leptospirosis in acutely febrile patients to decrease morbidity and mortality associated with this disease.

Scrub typhus: An under-reported and emerging threat - hospital based study from central and eastern Uttar Pradesh, India.

BACKGROUND & OBJECTIVES: Scrub typhus is a zoonotic rickettsial disease that is transmitted by the bite of the larval stage (chiggers) of trombiculid mites. The aim of this study was to determine the existence of scrub typhus in central and eastern Uttar Pradesh, India in patients with acute febrile illness (AFI) presenting to a super specialty tertiary level institute. METHODS: This prospective hospital-based study was conducted for a period of one year, from August 2018 to July 2019. About 2-5 mL of blood samples, along with clinical, epidemiological, and demographic data from a total of 125 patients presenting with acute febrile illness to outpatient and inpatient departments, were collected. ELISA testing tested the sera from blood samples for IgM antibodies against scrub typhus. Samples were also tested for dengue, leptospirosis, malaria and typhoid. RESULTS: During the study period, out of a total of 125 samples collected, 20% were found positive for IgM antibodies against scrub typhus. Demographically higher positivity was found in males, older age group, and in rural area. Rainfall was found to be important epidemiological parameter for presence of scrub typhus. Co-infection with dengue, leptospirosis and malaria was found. INTERPRETATION & CONCLUSION: Scrub typhus is found to be an important cause of acute febrile illness. It is necessary to include it in differential diagnosis of AFI cases even in absence of eschar. Diagnostic facilities of this as a screening test should be started in primary care centers or community health centers of rural areas of districts of central and eastern Uttar Pradesh, India.

Plasmid-Mediated AmpC (pAmpC) Genotypes Among Uropathogenic Escherichia coli: A Hospital-Based Study From Western Uttar Pradesh.

Introduction Resistance due to AmpC and extended-spectrum beta (ß)-lactamases (ESBLs) in Escherichia coli is an emerging problem worldwide. AmpC enzymes are a subclass of ß-lactamases that have a capacity to hydrolyze and deactivate a large range of ß-lactam antibiotics, particularly cephalosporins, penicillins, and monobactams, although frequently being susceptible to carbapenems and fourth-generation cephalosporins. The prevalence of plasmid-mediated AmpC (pAmpC) genotypes in uropathogenic E. coli isolates were looked at a tertiary care teaching hospital of Western Uttar Pradesh. Materials and methods A total of 312 non-repeat clinical E. coli isolates among patients presented with urinary tract infections (UTIs) were investigated by standard microbiological methods. Isolates were screened for the presence of ampC using a cefoxitin (30 µg) disc and confirmed using an inhibitor-based assay. Using multiplex polymerase chain reaction (PCR), six AmpC genotypes, namely, CIT, DHA, EBC, ACC, FOX, and MOX, were genotypically identified. Results A total of 152 (48.72%) uropathogenic E. coli isolates tested positive on the cefoxitin screening. Out of which, AmpC production was confirmed in 118/152 (77.63%) using a phenotypic method. In particular, the pAmpC gene was found in 56/152 (36.84%) isolates. CIT was the most common gene detected in this geographical area (57.14 %). Multiple genes, i.e., CIT and FOX, were also detected in 14.29% of the isolates.  Conclusion Identifying AmpC producers is important in routine microbiology laboratory as they are a nosocomial threat requiring strict adherence to infection control protocols. A confirmatory phenotypic test followed by genotypic tests will help in the correct and accurate identification of this resistance.

Evaluation of TaqMan based real-time PCR assay targeting LipL32 gene for leptospirosis in serologically positive human urine samples from north India.

PURPOSE: Rapid diagnosis of leptospirosis, through culture and/or serology, can be difficult without proper expertise and is often delayed because of the length of time required to obtain results. In India, especially in north India; leptospirosis is a grossly underreported disease, probably due to the lack of diagnostic modalities and lack of awareness of the disease among physicians. In this study, we aimed to identify leptospires in urine samples from serologically positive patients of leptospirosis through TaqMan based Real-time PCR using the LipL32 gene. Although environmental conditions are suitable in north India nowadays, very few studies were there and none by real-time PCR. METHOD: A total of 50 urine and blood samples (40 seropositive and 10 seronegative by IgM ELISA) were recruited in the study. Urine samples were tested by real-time PCR to detect Leptospira DNA and blood samples were used for microscopic agglutination test (MAT) evaluation. RESULT: We were able to detect leptospires in 20% (10/50) cases using our real-time PCR assay. In all 10 PCR positive urine samples, the LipL32 gene was detected and the lower limit of detection in L. interrogans DNA was found to be 20 GE/µL with a 95% cutoff value. CONCLUSION: We conclude that this real-time PCR assay with high specificity and sensitivity less prone to contamination may prove to be a promising approach for diagnosing acute leptospires in urine samples in the second week of acute febrile illness due to leptospiruria starts approximately in the second week of illness.