https://digitallibrary.bldedu.ac.in/xmlui/handle/123456789/1935 2026-04-09T12:57:52Z 2026-04-09T12:57:52Z Muktayakka, G https://digitallibrary.bldedu.ac.in/xmlui/handle/123456789/4165 2022-03-04T11:42:00Z 2021-09-01T00:00:00Z

Title: Cytokine Analysis And Drug Resistance Associated Genetic Polymorphism In Plasmodium Vivax Authors: Muktayakka, G Abstract: Introduction: Malaria is a vector born disease of major public health concern in several tropical and subtropical countries. Five different plasmodium species are known to cause malaria. For optimal public health measures, region specific prevalence of plasmodium species should be identified by optimal diagnostic methods available. Inflammatory cytokines play an important role in human immune responses to malaria. There should be cytokine balance between pro-inflammatory and anti-inflammatory cytokines. If there is dysregulation, amongst these pro-inflammatory and anti-inflammatory cytokines, it will lead to pathogenic effects. Major obstacle in the malaria prevention and eradication is the emergence of resistance in parasites towards many anti-malarial drugs. This significantly compromise the strategies used in controlling the infection. Aims and Objectives To study the prevalence of P.vivax and P.falciparum infections among suspected malaria cases and to analyze pro-inflammatory and anti-inflammatory cytokines implicated in malaria such as TNF-α, IFN-γ and IL-10, and TGF-β and to identify the mutation in drug resistance genes; pvmrd1 and pvdhfr of Plasmodium vivax clinical isolates to understand drug resistance pattern. Material and methods: A cross sectional study was conducted in 600 clinically suspected malaria cases. All the blood samples were screened by conventional PBS microscopy and rapid diagnostic tests (RDT). Blood samples positive for malaria were subjected to detection of cytokines TNF-α, IFN-γ, IL-10, and TGF-β by ELISA. Molecular confirmation of P.vivax and detection of pvmdr-1 gene and pvdfhr gene responsible for drug resistance in P.vivax were analysed. . Statistical analysis: Data will be analysed using SPSS software (version 20). The percentage analysis of the data will be given.Results: A total of 600 blood samples of malaria symptomatic cases were screened. 45 samles were found to be positive for malaria by microscopic observation and 51 samples by antigen detection by RDTs and 36 samples by PCR. Out of these 45 positive cases 33 (73%) were caused by P. vivax, 10 (22.2%) by P. falciparum and 2 (4.4%) were of mixed infection (P. vivax + P. falciparum) cases. Both the selected pro-inflammatory (TNF-α and IFN-γ) and anti-inflammatory (IL-10 and TGF-β) markers in the present study were found to be significantly elevated in malaria cases compared to healthy controls. In this study most of the P.vivax isolates had mutations in T958M (94.4%) & F1076L (83.3%) was observed and one isolate had mutations in Y976F (2.7%) pvmdr1 and wild type single type mutation in S58R and S117N amino acid positions of pvdhfr genes. Conclusion: The present study detected the presence of SNPs in both pvmdr-1 and pvdhfr gene in the selected geographical area. The frequency of mutations in these genes does not indicate the development of complete resistance to chloroquine and sulfadoxine-pyrimethamine in P. vivax. However, few SNPs detected in both genes suggested the probable early phase of resistance development.

2021-09-01T00:00:00Z Pramod, Manthalkar S https://digitallibrary.bldedu.ac.in/xmlui/handle/123456789/2911 2020-10-06T10:28:22Z 2020-01-01T00:00:00Z

Title: Serological & Molecular Characterization of Dengue Virus in a Tertiary Care Hospital of North Karnataka. Authors: Pramod, Manthalkar S Abstract: Dengue is a viral disease, caused by Flavivirus, transmitted by mosquito vector. Dengue virus infection may vary from mild Dengue fever to severe complications like, DHF and DSS. Dengue fever is the major cause of high mortality and morbidity in Tropical and Subtropical regions. 50 to 100 million people are affected every year globally. Dengue virus is a RNA virus and has got four different serotypes. Dengue presents with Fever, Arthralgia, Retro-orbital pain, Myalgia and in severe cases it may present with Nausea, Vomiting and bleeding tendency. Severity in dengue infection is because of the partial protection against the other serotypes and in multiple serotype infection the antibody dependent enhancement of the infection is seen. Laboratory diagnosis plays an important role in diagnosis of disease, which may help in proper treatment and immediate action, as there is no treatment available and vaccines are still under trial. There are various methods for detection of the Antigen and Antibodies of dengue virus in patient’s serum, which may help in diagnosis of the disease. This study was aimed to detect the incidence of the Dengue virus infection and to evaluate the proper dengue diagnostic test and to know the incidence of multiple serotype infection. Material and Methods: 1000 Dengue suspected patients as per WHO criteria were considered, serum samples were collected from the patients and subjected for the detection of Antigen and antibodies by rapid and ELISA method, all the NS-1 positive serum samples were subjected for detection of the viral RNA, serotyping was done to know the serotype present in the serum.Result: Out of 1000 samples, 462 serum samples were positive for Ag or Ab or both by rapid or ELISA. All the serum samples were subjected for testing by Rapid card method and ELISA. Male patients were more infected compared to women but significant difference was not observed. Age groups of 15-30 years were more infected followed by 30-45 age groups. More number of Dengue cases were seen in the month of October i.e. Post monsoon months, cases started in the month of June and ended in December. More number of cases was from Bidar city followed by Humnabad, Bhalki, Aurad and Baswakalyan. All the patients were suffering from fever, followed by headache, myalgia, body ache, arthralgia, nausea, vomiting and abdominal pain. All the NS-1 positive samples were subjected for RT PCR for the detection of the Dengue virus serotype. DENV-1, 2 and DENV3 were identified from the samples and detailed clinical correlation was done. Conclusion: ELISA is the most useful tool for detection of Dengue virus antigen or antibodies from patient’s serum. DENV-1, DENV-2 and DENV-3 are circulating with changing trend of serotype was observed, with severe clinical features in DENV-2 and 3 serotype infection. Serotype-1 showed unique conserved site Nco1.

2020-01-01T00:00:00Z Shriharsha, Hegde M.L https://digitallibrary.bldedu.ac.in/xmlui/handle/123456789/2910 2020-10-06T09:27:02Z 2020-01-01T00:00:00Z

Title: Serodiagnosis And Molecular Characterization Of Rickettsia In And Around Vijayapura North Karnataka, India. Authors: Shriharsha, Hegde M.L Abstract: Background Rickettsiosis is the most covert, re-emerging vector borne bacterial disease of public health importance distributed in many parts of the world. The disease is mostly go unnoticed or misdiagnosed due to low manifestation and non availability of specific diagnostic tests at all levels of healthcare setup. Failure of timely diagnosis and treatment leads to significant morbidity and mortality. Aim & Objectives: The present study aimed to demonstrate the presence of rickettsial infections in and around Vijayapur district using both serological and molecular diagnostic tests. Methods: In the present study, 572 blood samples from clinically suspected patients were screened for the presence of antibodies against rickettsial infections by routinely used Weil felix test. For the serological confirmation, all the serum samples were tested further by more sensitive and specific diagnostic assay, R.conorii and Scrub typhus specific IgM ELISA. To detect the etiological agent of the rickettsial infection, blood clot of ELISA positive samples were tested by nested PCR using the primers specific to the gene encoding different antigens of Rickettsia and Orientia. Sanger sequencing was performed to confirm the amplification of the gene. Phylogenetic tree was constructed to compare the sequences with other strains. Results: Out of 572 cases, 213 (37.23%) samples have shown titre of 1:160 and above with Weil felix OX antigens suggestive of seropositivity for rickettsial infection. Among 213 WF positive cases, 180 (31.46) samples have shown agglutination with OX2 antigen suggestive of SFG Rickettsial infection, and 63 (11.01%) cases shown agglutination with OXK antigen suggestive of Scrub typhus infection. In R. conorii IgM ELISA, 56 sampleswere positive out of 572. In Scrub typhus IgM ELISA, out of 432 samples 23 samples proved to be scrub typhus infection serologically. In nPCR for Scrub typhus, two samples have amplified successfully with the primers specific to 483bp segment of gene encoding 56kDa antigen of Orientia tsutsugumushi. Both the samples were sequenced by using Sanger sequencing technique. Sequences of the study sample have shown closest homology with many other strains reported from various parts of India and Southeast Asia. Phylogenetic analysis O. tsutsugumushi with sequences of the study have shown close relatedness with the strain reported from Korea. Several attempts were made to standardize the nested PCR for the detection of Rickettsia genus specific citrate synthase gene (gltA) and SFGR specific outer membrane protein gene (ompA) using specific primers. However, in spite of multiple attempts we could not standardize the PCR. Conclusion: Findings of our study demonstrated that Rickettsial infection is also circulating and causing acute febrile illness in and around Vijayapura, North Karnataka region. Significant seropositivity was observed for SFGR than Scrub typhus; however molecular detection of scrub typhus has proved its existence. More seropositivity was observed during the cooler months (August to January). Clinical diagnosis of rickettsial infection is really challenging as most of the common symptoms are non specific, and leads to misdiagnosis if not thoroughly examined. As the routinely used Weil felix test is lacking both sensitivity and specificity, inclusion of more specific tests like IgM ELISA and/or nPCR in routine diagnostic course would be highly beneficial for accurate diagnosis and patient management.

2020-01-01T00:00:00Z Sudheendra, Kulkarni https://digitallibrary.bldedu.ac.in/xmlui/handle/123456789/2473 2020-05-30T14:58:46Z 2019-09-01T00:00:00Z

Title: Biofilm Formation In Uropathogenic Escherichia Coli Strains; Relationship With Virulence Factors And Antimicrobial Resistance In Tertiary Care Hospital In North Karnataka Region Authors: Sudheendra, Kulkarni Abstract: Background: Urinary tract infection (UTI) is a clinical condition, with characteristic symptoms, colonization and multiplication of bacteria in significant numbers i.e., 105 cfu/ml within the urinary tract. At this juncture, antimicrobial resistance (AMR) in uropathogens has become one of the major concerns globally. Aim & Objectives: The present study aimed to demonstrate the biofilm formation in Uropathogenic Escherichia coli strains; characterize the phenotypic & genotypic virulence factors and their relationship with antimicrobial resistance. Methods: In the present study, 1000 suspected UTI cases were included. Urine samples were processed for culture and antimicrobial drug susceptibility testing. Only Escherichia coli (E. coli) isolates were further studied for production of biofilm, ESBL and haemolysin enzymes by phenotypic methods. Virulence genes, papEF, traT & PAI were detected by multiplex PCR. Molecular confirmation of virulence genes was done by Sanger sequencing. Sequences were studied for characterization and mutations. Results: From the 1000 urine samples, 395 E. coli were isolated. Higher resistance was observed with antibiotics - ampicillin (82.53%), cefuroxime (72.41%), amoxicillinclavulanic acid (71.90%), ceftriaxone (66.58%), ciprofloxacin (65.82%) and cefepime (57.47%). Further, in-vitro biofilm assay confirmed formation of biofilms by 71.39% isolates. Biofilm-forming E. coli strains developed higher degree of resistance towards antibiotics ampicillin (87.36%) followed by cefuroxime (81.58%), amoxicillin-clavulanic acid (77.61%), ciprofloxacin (71.48%), cefepime (64.98%) and ceftriaxone (54.6%). Phenotypic methods detected 62.3% isolates as ESBL producers and 40.2% were β-haemolytic. Virulence gene characterization revealed presence of gene traT in 73.2% isolates, PAI in 62.9% isolates and papEF among 33.5% isolates. ESBL & Hemolysin producing UPEC exhibited significantly higher resistant to ampicillin, amoxicillin-clavulanic acid, aztreonam, cefriaxone, cefuroxime, cefepime, ciprofloxacin, chloramphenicol, gentamicin and norfloxacin. Association between the expression of virulence genes and antimicrobial resistance was observed. Statistically proved association was seen with expression of genes RPAI and traT with antibioticsnitrofurantoin and amikacin respectively. All the isolates were sensitive to antibiotics nitrofurantoin, piperacillin-tazobactam, and imipenem. Strains showed 99% sequence identity in Sanger sequencing and no mutations were detected among the study strains. Conclusion: Increase in trend of resistance was observed with antibiotics which were routinely used to treat UTI. Predominance of the traT gene and PAI markers was seen among UPEC strains. Results indicated significant correlation between phenotypic and genotypic virulence factors and antibiotic resistance. Antibiotics nitrofurantoin, piperacillin - tazobactam, and imipenem can be effective for severe UTIs. This study concludes that expression of virulence factors by UPEC strains is responsible for increased antibiotic resistance. Hence, characterizing the UPEC strains help clinicians and microbiologists to reach a better therapeutic outcomes and treatment regimens in this region.

2019-09-01T00:00:00Z