Detection of adeB and basD Genes in Lung Tissue Samples of Rattus Norvegicus Injected with Acinetobacter Baumannii
DOI:
https://doi.org/10.37287/ijghr.v8i2.752Keywords:
acinetobacter baumanni, adeB gene, basD gene, lung tissue, PCR, rattus norvegicusAbstract
Acinetobacter baumannii is an opportunistic pathogen and a major cause of nosocomial infections, characterized by high resistance to multiple antibiotics. The success of its colonization and virulence is influenced by resistance factors such as the adeB gene and nutrient acquisition factors such as the basD gene. This study aimed to detect the presence of adeB and basD genes in lung tissue of Rattus norvegicus intravenously injected with clinical isolates of Acinetobacter baumannii. Six healthy female rats were divided into treatment groups with observation periods of 10 hours and 18 hours post-infection. Lung tissues were collected for DNA extraction, followed by PCR amplification using specific primers for adeB (549 bp) and basD (533 bp). Gel electrophoresis results showed that both genes were clearly detected in the bacterial isolate positive control, but not in lung tissue samples post-infection. The failure of detection in tissue samples is likely influenced by the low bacterial load, host-dependent regulation of gene expression, and the presence of PCR inhibitors derived from lung tissue. These findings indicate that although adeB and basD play crucial roles in resistance and virulence, their presence in host tissues is not always detectable by conventional PCR. Therefore, more sensitive detection methods are required for molecular analysis in in vivo models.
References
Ahmad, I., Karah, N., Nadeem, A., Wai, S. N., & Uhlin, B. E. (2019). Analysis of colony phase variation switch in Acinetobacter baumannii clinical isolates. PLOS ONE, 14(1), e0210082. https://doi.org/10.1371/journal.pone.0210082
Akhmad, A. F., Ulfa, M., & Azuma, M. (2024). Antibiotic Susceptibility Patterns among Indonesian Adults Hospitalized with Pneumonia. Jurnal Respirasi, 10(1), 6–13. https://doi.org/10.20473/jr.v10-I.1.2024.6-13
Almaghrabi, M. K., Joseph, M. R. P., Assiry, M. M., & Hamid, M. E. (2018). Multidrug-Resistant Acinetobacter baumannii : An Emerging Health Threat in Aseer Region, Kingdom of Saudi Arabia. Canadian Journal of Infectious Diseases and Medical Microbiology, 2018, 1–4. https://doi.org/10.1155/2018/9182747
Anduni, L., Molina, H., Zazueta, A., Cancino, J., Ponce, C., Chakoory, O., Comtet-Marre, S., Tapia, C. V., Peyret, P., Gotteland, M., & Magne, F. (2024). Optimization of lung tissue pre-treatment by bead homogenization for subsequent culturomics. Scientific Reports, 14(1), 22724. https://doi.org/10.1038/s41598-024-69736-2
Aulia, N., Ahda, Y., Achyar, A., & Putri, D. H. (2023). Desain Primer dan Optimasi Suhu Annealing untuk Amplifikasi Gen RET. BIOSEL (Biology Science and Education): Jurnal Penelitian Science dan Pendidikan, 12(1), 70-77.
Ayoub Moubareck, C., & Hammoudi Halat, D. (2020). Insights into Acinetobacter baumannii: A Review of Microbiological, Virulence, and Resistance Traits in a Threatening Nosocomial Pathogen. Antibiotics, 9(3), 119. https://doi.org/10.3390/antibiotics9030119
Breslow, J. M., Meissler, J. J., Hartzell, R. R., Spence, P. B., Truant, A., Gaughan, J., & Eisenstein, T. K. (2011). Innate Immune Responses to Systemic Acinetobacter baumannii Infection in Mice: Neutrophils, but Not Interleukin-17, Mediate Host Resistance. Infection and Immunity, 79(8), 3317–3327. https://doi.org/10.1128/IAI.00069-11
Byun, J.-H., Park, S. E., Seo, M., Jang, J., Hwang, M. S., Song, J. Y., Chang, C. L., & Kim, Y. A. (2021). Controlling an Outbreak of Multidrug-resistant Acinetobacter baumannii in a Pediatric Intensive Care Unit: a Retrospective Analysis. Journal of Korean Medical Science, 36(46). https://doi.org/10.3346/jkms.2021.36.e307
Conde-Pérez, K., Vázquez-Ucha, J. C., Álvarez-Fraga, L., Ageitos, L., Rumbo-Feal, S., Martínez-Guitián, M., Trigo-Tasende, N., Rodríguez, J., Bou, G., Jiménez, C., Beceiro, A., & Poza, M. (2021). In-Depth Analysis of the Role of the Acinetobactin Cluster in the Virulence of Acinetobacter baumannii. Frontiers in Microbiology, 12. https://doi.org/10.3389/fmicb.2021.752070
Delaplace, M., Huet, H., Gambino, A., & Le Poder, S. (2021). Feline Coronavirus Antivirals: A Review. Pathogens, 10(9), 1150. https://doi.org/10.3390/pathogens10091150
Dewanata, P. A., & Mushlih, M. (2021), Differences in DNA Purity Test Using UV-Vis Spectrophotometer and Nanodrop Spectrophotometer in Type ype 2 Diabetes Mellitus Patients. Indonesian Journal of Innovation Srubes, 15. https://doi.org/10.21070/ijins. v151.553
Hasan, T., Choi, C. H., & Oh, M. H. (2015). Genes Involved in the Biosynthesis and Transport of Acinetobactin in Acinetobacter baumannii. Genomics & Informatics, 13(1), 2. https://doi.org/10.5808/GI.2015.13.1.2
Huang, K., Zhang, J., Li, J., Qiu, H., Wei, L., Yang, Y., & Wang, C. (2024). Exploring the Impact of Primer–Template Mismatches on PCR Performance of DNA Polymerases Varying in Proofreading Activity. Genes, 15(2), 215. https://doi.org/10.3390/genes15020215
Jakobsen, T. H., Kirkegaard, J. B., Lichtenberg, M., Kvich, L. A., Gottlieb, H., McNally, M., & Bjarnsholt, T. (2025). Detection limitations of bacteria in tissue samples. Bone & Joint Research, 14(6), 560–567. https://doi.org/10.1302/2046-3758.146.BJR-2024-0410.R1
Jansriphibul, K., Krohn, C., & Ball, A. S. (2025). Sources of variability for viability PCR using propidium monoazide. Microbiological Research, 128224. https://doi.org/10.1016/j.micres.2025.128224
Kim, M., Kim, D. Y., Song, W. Y., Park, S. E., Harrison, S. A., Chazin, W. J., Oh, M. H., & Kim, H. J. (2021). Distinctive Roles of Two Acinetobactin Isomers in Challenging Host Nutritional Immunity. MBio, 12(5). https://doi.org/10.1128/mBio.02248-21
Kusumawati, S. D., Hardianto. 1., Nurlatifah, Pracoyo, A. A., & Handayani, N. A. (2023). Perbandingan Nilai Pengukuran Kuantitatif Isolat Asam Ribonukleat (RNA) Menggunakan Spektrofotometer Nanodrop dan Mikrodrop pada Sampel Hepar Ayam (Gallus gallus domesticus). Indonesian Jurnal of Laboratory. 62-71
Latham S, Hughes E, Budgen B, Morley A (2023) Inhibition of the PCR by genomic DNA. PLoS ONE 18(4): e0284538. https://doi.org/10.1371/journal.pone.0284538
Li, S., Wei, B., Xu, L., Cong, C., Murtaza, B., Wang, L., Li, X., Li, J., Xu, M., Yin, J., & Xu, Y. (2024). A novel Saclayvirus Acinetobacter baumannii phage genomic analysis and effectiveness in preventing pneumonia. Applied Microbiology and Biotechnology, 108(1), 428. https://doi.org/10.1007/s00253-024-13208-0
Lu, X., Lafon, O., Trébosc, J., & Amoureux, J.-P. (2012). Detailed analysis of the S-RESPDOR solid-state NMR method for inter-nuclear distance measurement between spin-1/2 and quadrupolar nuclei. Journal of Magnetic Resonance, 215, 34–49. https://doi.org/10.1016/j.jmr.2011.12.009
Nageeb, W. M., AlHarbi, N., Alrehaili, A. A., Zakai, S. A., Elfadadny, A., & Hetta, H. F. (2023). Global genomic epidemiology of chromosomally mediated non-enzymatic carbapenem resistance in Acinetobacter baumannii: on the way to predict and modify resistance. Frontiers in Microbiology, 14. https://doi.org/10.3389/fmicb.2023.1271733
Nazarov, P. A., Kuznetsova, A. M., & Karakozova, M. V. (2022). Multidrug Resistance Pumps as a Keystone of Bacterial Resistance. Moscow University Biological Sciences Bulletin, 77(4), 193–200. https://doi.org/10.3103/S009639252204006X
Nwe, M.K., Jangpromma, N. & Taemaitree, L. Evaluation of molecular inhibitors of loop-mediated isothermal amplification (LAMP). Sci Rep 14, 5916 (2024). https://doi.org/10.1038/s41598-024-55241-z
Q, H., Y, L., S, Y., & D, H. (2014). The effect of six common PCR inhibitors on DNA polymerase and DNA template. International Journal of Forensic Science & Pathology, 61–64. https://doi.org/10.19070/2332-287X-1400017
Rafiei, E., Shahini Shams Abadi, M., Zamanzad, B., & Gholipour, A. (2022). The frequency of efflux pump genes expression in Acinetobacter baumannii isolates from pulmonary secretions. AMB Express, 12(1), 103. https://doi.org/10.1186/s13568-022-01444-4
Revathy, S., Sabu, K. K., & Anilkumar, S. (2025). Comparative study of DNA and RNA extraction methods for high-quality nucleic acid isolation from Cullenia exarillata A. Robyns. Cellular and Molecular Biology, 71(6), 54-61. https://doi.org/10.14715/cmb/2025.71.6.7
Reyes, C., Sajó, Z., Lucas, M. S., Sinha, A., Schwarze, F. W. M. R., Ribera, J., & Nyström, G. (2022). Cocultivation of White-Rot Fungi and Microalgae in the Presence of Nanocellulose. Microbiology Spectrum, 10(5). https://doi.org/10.1128/spectrum.03041-22
Suresh, G., Lodha, T. D., Indu, B., Sasikala, C., & Ramana, C. V. (2019). Taxogenomics Resolves Conflict in the Genus Rhodobacter: A Two and Half Decades Pending Thought to Reclassify the Genus Rhodobacter. Frontiers in Microbiology, 10. https://doi.org/10.3389/fmicb.2019.02480
Tansho-Nagakawa, S., SATO, Y., UBAGAI, T., KIKUCHI-UEDA, T., KAMOSHIDA, G., NISHIDA, S., & ONO, Y. (2021). Histopathological Analysis of Acinetobacter baumannii Lung Infection in a Mouse Model. Polish Journal of Microbiology, 70(4), 469–477. https://doi.org/10.33073/pjm-2021-044
Wadsworth, P. (2012). Using Cultured Mammalian Cells to Study Mitosis. Cold Spring Harbor Protocols, 2012(2), pdb.ip067850. https://doi.org/10.1101/pdb.ip067850
Wang, H., Xu, Q., Heng, H., Zhao, W., Ni, H., Chen, K., Wai Chan, B. K., Tang, Y., Xie, M., Peng, M., Chi Chan, E. W., Yang, G., & Chen, S. (2024). High mortality of Acinetobacter baumannii infection is attributed to macrophage-mediated induction of cytokine storm but preventable by naproxen. EBioMedicine, 108, 105340. https://doi.org/10.1016/j.ebiom.2024.105340
Xu, C. F., Bilya, S. R., & Xu, W. (2019). adeABC efflux gene in Acinetobacter baumannii. New Microbes and New Infections, 30, 100549. https://doi.org/10.1016/j.nmni.2019.100549
Yoon, E.-J., Balloy, V., Fiette, L., Chignard, M., Courvalin, P., & Grillot-Courvalin, C. (2016). Contribution of the Ade Resistance-Nodulation-Cell Division-Type Efflux Pumps to Fitness and Pathogenesis of Acinetobacter baumannii. MBio, 7(3). https://doi.org/10.1128/mBio.00697-16
Zhang, W. Z., Butler, J. J., & Cloonan, S. M. (2019). Smoking-induced iron dysregulation in the lung. Free Radical Biology and Medicine, 133(10), 238–247. https://doi.org/10.1016/j.freeradbiomed.2018.07.024
Zhao, A., Sun, J., & Liu, Y. (2023). Understanding bacterial biofilms: From definition to treatment strategies. Frontiers in Cellular and Infection Microbiology, 13. https://doi.org/10.3389/fcimb.2023.1137947
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