Helicobacter pylori izolasyonu ve Cag A ve Vac A virulens genler temelinde izolatların genotiplendirilmesi

Year 2026, Volume: 15 Issue: 1, 55 - 63, 27.03.2026

Özden Dellal , Sevil Erdenlig Gürbilek

https://doi.org/10.31196/huvfd.1890481

https://izlik.org/JA59CK22JK

Abstract

Bu çalışmada, gastroduodenal şikayetleri olan hastaların mide biyopsilerinden H. pylori suşlarını izole etmek ve bu suşları genotiplendirmek amaçlandı. İzolasyon oranımız %32,8 (64/21) idi, bu da H. pylori enfeksiyonunun tanısında kültürün nispeten zayıf bir araç olduğunu göstermektedir. CagA, VacA s1/s2, VacA m1/m2 genotiplerinin tanımlanması için multipleks PCR kullandık. Sonuçlarımız, suşların %71,4'ünün hem CagA pozitif hem de CagA negatif olduğunu, en yaygın VacA genotipinin s1/m1 (%100) olduğunu ve bunu s1/m2 genotipinin (%62,5) izlediğini gösterdi. Sadece bir izolat s2/m2 idi, bu da numunelerin klinik semptomları olan hastalardan alındığı için beklenen bir bulguydu. Allelik VacA kombinasyonları açısından, Cag A+ numunelerde, 9 (%42,8), 5 (%23,8) ve 1 (%4,8) suş, sırasıyla s1/m1, s1/m2, s2/m2 genotiplerini taşıyordu. Cag A- suşlarının hiçbirinde s1/m1 VacA genotipi saptanmamıştır. Ancak, s1/m2 ve s2/m2 VacA genotipleri bu grupta eşit oranda (%14,3) bulunmuştur. s1/m1 VacA genotipi ile CagA durumu arasındaki ilişki istatistiksel olarak anlamlı bulunmuştur (P < 0,05).

Keywords

Helicobacter pylori , genotiplendirme , vagA/cagA

Ethical Statement

Written consent was taken from human patients. Before starting the study, approval was taken from Harran University Clinical Research Ethical Board Committee (HRU/24.19.50).

Supporting Institution

yok

Thanks

Çalışma materyallerinin temini ve makalenin hazırlanmasına sağladığı değerli katkılar için Prof. Dr. Savaş Cumali Efe’ye teşekkür ederiz.

Isolation of Helicobacter pylori and Genotyping the Isolates Based on cagA and vacA Virulence Genes

https://izlik.org/JA59CK22JK

Abstract

In this study, we aimed to isolate and genotype H. pylori strains from gastric biopsies of patients having gastroduodenal complaints. Our isolation rate was 32.8% (64/21),
suggesting that culture was a relatively weak diagnostic tool for H. pylori infection. We used multiplex PCR to identify the cagA, vacA s1/s2, and vacA m1/m2 genotypes. Our results indicated that 71.4% of the strains were both cagA-positive and cagA-negative. The predominant vacA genotype was s1/m1 (100%), followed by s1/m2 (62.5%). Only one isolate was s2/m2, as expected, since samples were obtained from patients with clinical symptoms. In terms of allelic vacA combinations, in cagA+ samples, 9 (42.8%), 5 (23.8%), and 1(4.8%) strains carried s1/m1, s1/m2, and s2/m2 genotypes, respectively. None of the cagA- strain was determined to have s1/m1 vacA genotype. However, s1/ m2 and s2/m2 vacA genotypes were found equally (14.3%) in this group. The association between s1/m1 vacA genotype and cagA status was found statistically significant (P<0.05).

Keywords

Genotyping , Helicobacter pylori , vagA/cagA.

Ethical Statement

Ethical approval was granted by the Harran University Clinical Research Ethics Committee: (Session No: 19, Approval No: HRU/24.19.50, Date: 02 December 2024).

Supporting Institution

No

Thanks

We would like to thank to Prof. Dr. Savaş Cumali Efe for valuable contributions to the procurement to study metarials and the preparation of the manuscript

References

• Almagro-Martínez, C., Alenda-Botella, A., Botella-Juan, L. (2025). Systematic review on the zoonotic potential of Helicobacter pylori. Discover Public Health, 22(1), 432. https://doi.org/10.1186/s12982-025-00834-w
• Atherton, J. C., Cao, P., Peek, R. M., Tummuru, M. K., Blaser, M. J., Cover, T. L. (1995). Mosaicism in vacuolating cytotoxin alleles of Helicobacter pylori: association of specific vacA types with cytotoxin production and peptic ulceration. Journal of Biological Chemistry, 270(30), 17771-17777. https://doi.org/10.1074/jbc.270.30.17771
• Aydin, F. A. R. U. K., Kaklikkaya, N. E. Ş. E., Ozgur, O. R. H. A. N., Cubukcu, K., Kilic, A. O., Tosun, I., & Erturk, M. (2004). Distribution of vacA alleles and cagA status of Helicobacter pylori in peptic ulcer disease and non-ulcer dyspepsia. Clinical microbiology and infection, 10(12), 1102-1104. https://doi.org/10.1111/j.1469-0691.2004.00989.x
• Blaser, M. J., Chyou, P. H., Nomura, A. (1995). Age at establishment of Helicobacter pylori infection and gastric carcinoma, gastric ulcer, and duodenal ulcer risk. Cancer research, 55(3), 562-565.
• Bilman, F. B., Özdemir, M., Baysal, B., Kurtoğlu, M. G. (2016). Prevalence of H. pylori in gastric biopsy specimen in the southeastern region of Turkey. The Journal of Infection in Developing Countries, 10(11), 1177-1182. https://doi.org/10.3855/jidc.6690
• Chattopadhyay, S., Patra, R., Ramamurthy, T., Chowdhury, A., Santra , A., Dhali, G. K., & Mukhopadhyay, A. K. (2004). Multiplex PCR assay for rapid detection and genotyping of Helicobacter pylori directly from biopsy specimens. Journal of clinical microbiology, 42(6), 2821-2824. https://doi.org/10.1128/jcm.42.6.2821-2824.2004
• Chen, Y. C., Malfertheiner, P., Yu, H. T., Kuo, C. L., Chang, Y. Y., Meng, F. T., & Liou, J. M. (2024). Global prevalence of Helicobacter pylori infection and incidence of gastric cancer between 1980 and 2022. Gastroenterology, 166(4), 605-619. https://doi.org/10.1053/j.gastro.2023.12.022
• Chiurillo, M. A., Moran, Y., Canas, M., Valderrama, E., Granda, N., Sayegh, M., Ramírez, J. L. (2013). Genotyping of Helicobacter pylori virulence-associated genes shows high diversity of strains infecting patients in western Venezuela. International Journal of Infectious Diseases, 17(9), e750-e756. https://doi.org/10.1016/j.ijid.2013.03.004
• Erdoğdu, C., Saribaş, Z., Yilmaz, Y. A. (2014). Detection of cagA and vacA genotypes of Helicobacter pylori isolates from a university hospital in Ankara region, Turkey. Turkish journal of medical sciences, 44(1), 126-132. https://doi.org/10.3906/sag-1209-72
• Erzin, Y., Koksal, V., Altun, S., Dobrucali, A. H. M. E. T., Aslan, M., Erdamar, S., & Kocazeybek, B. (2006). Prevalence of Helicobacter pylori vacA, cagA, cagE, iceA, babA2 genotypes and correlation with clinical outcome in Turkish patients with dyspepsia. Helicobacter, 11(6), 574-580. https://doi.org/10.1111/j.1523-5378.2006.00461.x
• Germani, Y., Dauga, C., Duval, P., Huerre, M., Levy, M., Pialoux, G., Grimont, P. A. D. (1997). Strategy for the detection of Helicobacter species by amplification of 16S rRNA genes and identification of H. felis in a human gastric biopsy. Research in microbiology, 148(4), 315-326. https://doi.org/10.1016/S0923-2508(97)81587-2
• Ragab, R., Abdel Hamid, H. S., Emara, N., Metwally, M., El-Kholy, H. R. (2022). Patient and Microbial Factors Affecting Culture of Helicobacter Pylori. Benha medical journal, 39(1), 309-318. https://doi.org10.21608/BMFJ.2022.109333.1518
• Jones, K. R., Whitmire, J. M., Merrell, D. S. (2010). A tale of two toxins: Helicobacter pylori CagA and VacA modulate host pathways that impact disease. Frontiers in microbiology, 1, 115. https://doi.org/10.3389/fmicb.2010.00115
• Kaya, A. D., Öztürk, C. E., Akcan, Y., Behçet, M., Karakoç, A. E., Yücel, M., & Tuncer, S. (2007). Prevalence of Helicobacter pylori in symptomatic patients and detection of clarithromycin resistance using melting curve analysis. Current therapeutic research, 68(3), 151-160. https://doi.org/10.1016/j.curtheres.2007.06.001
• Hamze, H., Payne, M., Stefanovic, A., Lowe, C. F., Romney, M. G., Matic, N. (2025). Helicobacter pylori culture positivity and antimicrobial susceptibility profiles (Vancouver, Canada). Journal of Antimicrobial Chemotherapy, 80(6), 1640-1646. https://doi.org/10.1093/jac/dkaf114
• Hatakeyama, M. (2006). Helicobacter pylori CagA—a bacterial intruder conspiring gastric carcinogenesis. International journal of cancer, 119(6), 1217-1223. https://doi.org/10.1002/ijc.21831
• Nagiyev, T., Yula, E., Abayli, B., Koksal, F. (2009). Prevalence and genotypes of Helicobacter pylori in gastric biopsy specimens from patients with gastroduodenal pathologies in the Cukurova region of Turkey. Journal of clinical microbiology, 47(12), 4150-4153. https://doi.org/10.1128/jcm.00605-09
• Nguyen, T. M. N., Tran, V. H., Ha, T. M. T. (2024). Helicobacter pylori cagA, vacA, and iceA genotypes and clinical outcomes: a cross-sectional study in central Vietnam. Brazilian Journal of Microbiology, 55(2), 1393-1404. https://doi.org/10.1007/s42770-024-01328-8
• Ozbey, G., Dogan, Y., Demiroren, K., Ozercan, I. H. (2015). Prevalence of Helicobacter pylori in children in eastern Turkey and molecular typing of isolates. Brazilian Journal of Microbiology, 46, 505-511. https://doi.org/10.1590/S1517-838246220140234
• Paredes-Osses, E., Sáez, K., Sanhueza, E., Hebel, S., González, C., Briceño, C., García Cancino, A. (2017). Association between cagA, vacAi, and dupA genes of Helicobacter pylori and gastroduodenal pathologies in Chilean patients. Folia microbiologica, 62(5), 437-444. https://doi.org/10.1007/s12223-017-0514-y
• Peek Jr, R. M., Blaser, M. J. (2002). Helicobacter pylori and gastrointestinal tract adenocarcinomas. Nature Reviews Cancer, 2(1), 28-37. https://doi.org/10.1038/nrc703
• Perez-Perez, G. I., Rothenbacher, D., Brenner, H. (2004). Epidemiology of Helicobacter pylori infection. Helicobacter, 9, 1-6. https://doi.org/10.1111/j.1083-4389.2004.00248.x
• Perez-Perez, G. I., Van, T. N., Huong, D. T., Zhan, G., Anh, D. N., Nguyet, N. T., & Hong-Hanh, N. T. (2016). Isolation and characterization of Helicobacter pylori recovered from gastric biopsies under anaerobic conditions. Diagnostic microbiology and infectious disease, 86(2), 136-140. https://doi.org/10.1016/j.diagmicrobio.2016.07.009
• Rudi, J., Kolb, C., Maiwald, M., Kuck, D., Sieg, A., Galle, P. R., & Stremmel, W. (1998). Diversity of Helicobacter pylori vacA and cagA genes and relationship to VacA and CagA protein expression, cytotoxin production, and associated diseases. Journal of Clinical Microbiology, 36(4), 944-948. https://doi.org/10.1128/jcm.36.4.944-948.1998
• Shiota, S., Suzuki, R., Yamaoka, Y. (2013). The significance of virulence factors in H elicobacter pylori. Journal of digestive diseases, 14(7), 341-349. https://doi.org/10.1111/1751-2980.12054
• Yarahmadi, A., Afkhami, H. (2025). Potential relationship between Helicobacter pylori infection and autoimmune disorders: A narrative review. Microbial Pathogenesis, 205, 107572. https://doi.org/10.1016/j.micpath.2025.107572