Molecular Characterization of Multi-Drug Resistant Clinical Isolates from Healthcare Facilities

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DOI: 10.21522/TIJPH.2013.12.04.Art043

Authors : P. Sivagurunathan, Yagniyasree Manogaran, Pasiyappazham Ramasamy, M. Kiran Kumar, C. Uma, V. Praveen Kumar

Abstract:

This study aimed to analyze multi-drug resistant clinical isolates collected from healthcare facilities at the molecular level. Bacterial samples from a hospital were cultured on selective media to isolate Acinetobacter, E. coli, Klebsiella sp., and Pseudomonas sp., and characterized by Gram staining and biochemical tests. DNA was extracted from overnight cultures and used for PCR amplification of the V1-V3 and V3-V9 regions of the 16S rDNA. The PCR products were confirmed on agarose gels, purified, and sequenced. Sequences were analyzed using BLAST for species identification, and accession numbers were obtained from the NCBI GenBank database. Morphological and biochemical data for pathogenic bacteria, such as Acinetobacter (K1), E. coli (K2), Klebsiella (K3), and Pseudomonas sp. (K4) were analyzed. The 16s rDNA PCR products were sequenced using a commercial automated sequencer and were submitted to GenBank and NCBI. Bacterial sequencing analysis and comparison revealed that isolates K1, K2, K3, and K4 were Acinetobacter, E. coli, Klebsiella, and Pseudomonas sp. since they had a 100% homology with the library. The species-level identification of bacteria K1, K2, K3, and K4 is based on the highest identity (90%) and their GenBank accession codes are APK1, APK2, APK3, and APK4. This study suggests that samples from tertiary hospitals contain high loads of antibiotic-resistant bacteria. A change in the antibiotic sensitivity of the identified bacteria needs to be monitored, as there is limited availability of newer medicines, and the emergence of resistant bacteria far exceeds the rate of new antibiotic development.

References:

[1]    Hauhnar, L., Pachuau, L., Lalhruaitluanga, H., 2018, Isolation and characterization of multi-drug resistant bacteria from hospital wastewater sites around the city of Aizawl, Mizoram. Advances in Bioscience and Biotechnology, 9(07), 311-321.

[2]    Ravi, G. S., Ellappan, K., Narasimha, H. B., Rudresh, S. M., 2021, Isolation and Molecular Characterization of Multi Drug Resistant Gram-Negative Bacteria from Clinical Samples in Tertiary Care Hospital, South India. RGUHS Journal of Medical Sciences, 11(4).

[3]    Tang, S. S., Apisarnthanarak, A., Hsu, L. Y., 2014, Mechanisms of β-lactam antimicrobial resistance and epidemiology of major community-and healthcare-associated multidrug-resistant bacteria. Advanced Drug Delivery Reviews, 78, 3-13.

[4]    Khan, D., Zeb, M., Khattak, S. K., Shah, A. A., Abdullah, M., Bilal, M., 2023, Molecular characterization and antibiotic susceptibility pattern of bacterial strains isolated from diabetic foot ulcers. Endocrine and Metabolic Science, 12, 100136.

[5]    Momin, M. H. F. A., Liakopoulos, A., Phee, L. M., Wareham, D. W., 2017, Emergence and nosocomial spread of carbapenem-resistant OXA-232-producing Klebsiella pneumoniae in Brunei Darussalam. Journal of global Antimicrobial Resistance, 9, 96-99.

[6]    Kamala, K., Sivaperumal, P., 2023, Predominance of multi-drug resistant extended spectrum β lactamase producing bacteria from marine fishes. Environmental Pollution, 323, 121314.

[7]    Bhatia, A., Kalra, J., Kohli, S., Kakati, B., Kaushik, R., 2018, Antibiotic resistance pattern in intensive care unit of a tertiary care teaching hospital. International Journal of Basic and Clinical Pharmacology, 7(5), 906-911.

[8]    Masseron, A., Poirel, L., Ali, B. J., Syed, M. A., Nordmann, P., 2019, Molecular characterization of multidrug-resistance in Gram-negative bacteria from the Peshawar teaching hospital, Pakistan. New Microbes and New Infections, 32, 100605.

[9]    Sivaperumal, P., Kamala, K., Ganapathy, D. M., Dharani, G., Sundarrajan, S., Ramakrishna, S., 2023, Fabrication of AgNPs mediated fibrous membrane from Rhizophora mucronata mangrove plant extract for biological properties. Journal of Drug Delivery Science and Technology, 86, 104710.

[10]   Soni, M., Pitchiah, S., Suresh, V., Ramasamy, P., 2024, Fabrication and partial characterization of silver nanoparticles from Mangrove (Avicennia marina) Leaves and Their Antibacterial Efficacy Against Oral Bacteria. Cureus, 16(1).

[11]   Zarras, C., Karampatakis, T., Pappa, S., Iosifidis, E., Vagdatli, E., Roilides, E., Papa, A., 2023, Genetic characterization of carbapenem-resistant Klebsiella pneumoniae clinical isolates in a tertiary hospital in Greece, 2018–2022. Antibiotics, 12(6), 976.

[12]   Pauline, C. R., Akshita., Pavithra, T., Kannan, K., Sivaperumal, P., 2024, Characterization and biological activity of silver nanoparticles from (Rhizophora mucronata) Mangrove Extract. Nano LIFE, 2450018.

[13]   Choushette, B. B., Satpute, R. A., 2022, Isolation and characterization of multidrug resistance bacteria from hospital sewage samples, Maharashtra, India. African Journal of Biotechnology, 21(1), 16-25.

[14]   Ramasamy, P., Subhapradha, N., Srinivasan, A., Shanmugam, V., Krishnamoorthy, J., Shanmugam, A., 2011, In vitro evaluation of the antimicrobial activity of methanolic extract from selected species of cephalopods on clinical isolates. African Journal Microbiology Research, 5(23), 3884-3889.

[15]   Gupta, M., Naik, A. K., Singh, S. K., 2019, Bacteriological profile and antimicrobial resistance patterns of burn wound infections in a tertiary care hospital. Heliyon, 5(12).

[16]   Barai, L., Fatema, K., Haq, J. A., Faruq, M. O., Ahsan, A. A., Morshed, M. A. H. G., Hossain, M. B., 2010, Bacterial profile and their antimicrobial resistance pattern in an intensive care unit of a tertiary care hospital of Dhaka. Ibrahim Medical College Journal, 4(2), 66-69.

[17]   Kumar, A., Tanwar, S., Chetiwal, R., Kumar, R., 2022, Nosocomial infections-related antimicrobial resistance in a multidisciplinary intensive care unit. MGM Journal of Medical Sciences, 9(1), 12-18.

[18]   Chaudhary, L., Pandey, A., Singh, P., Chaturvedi, P., Bisht, A.S., 2023, Bacterial Profile and Antimicrobial susceptibility pattern of gram-negative bacteria isolated from skin and soft tissue infections in a tertiary care hospital of western Uttar Pradesh. www.ijphrd.Com, 14(3), 135.

[19]   Suma, P., Swetha, C. S., Sudhanthiramani., Goud, S. S., Annie Supriya., Jagadeesh Babu, A., 2016, A study on the antibiotic resistance patterns of Staphylococcus aureus isolated from market milk in and around Tirupati, Andhra Pradesh. International Journal of Recent Scientific Research, 7(4), 10429-10435.

[20]   Lagha, R., Abdallah, F. B., ALKhammash, A. A., Amor, N., Hassan, M. M., Mabrouk, I., Alhomrani, M., Gaber, A., 2021, Molecular characterization of multidrug-resistant Klebsiella pneumoniae clinical isolates recovered from King Abdulaziz Specialist Hospital at Taif City, Saudi Arabia. Journal of Infection and Public Health, 14(1), 143-151.

[21]   Zou, Z. Y., Lei, L., Chen, Q. Y., Wang, Y. Q., Cai, C., Li, W. Q., Zhang, Z., Shao, B., Wang, Y., 2019, Prevalence and dissemination risk of antimicrobial-resistant Enterobacteriaceae from shared bikes in Beijing, China. Environment International, 132, 105119.

[22]   Subhapradha, N., Ramasamy, P., Srinivasan, A., Shanmugam, A., Shanmugam, V., 2013, Preparation of chitosan derivatives from gladius of squid Sepioteuthis lessoniana (Lesson, 1830) and antimicrobial potential against human pathogens. Journal of Biological Sciences, 13(4), 257.

[23]   Zhang, Y., Li, D., Yang, Y., Su, J., Xu, X., Wang, M., Chen, Y., Li, Y., 2021, Clinical and molecular characteristics of Chryseobacterium indologenes isolates at a teaching hospital in Shanghai, China. Annals of Translational Medicine, 9(8).