International network for natural sciences – research journal
  • mendeley icon
  • linkedin icon
  • google plus icon
  • twitter icon
  • google scholar icon
  • facebook icon

PCR based identification of Helicobacter pylori infection in saliva samples

By: Komal Aman, Bashir Ahmad, Sajid Ali, Saad Ahmad Khan, Md Motiar Rahman, Farhana Jabeen Shah, Muslim Raza, Huang Linfang

Key Words: Helicobacter pylori, Saliva culture, Noninvasive, Polymerase chain reaction, Sequencing

Int. J. Biosci. 15(3), 496-504, September 2019.

DOI: http://dx.doi.org/10.12692/ijb/15.3.496-504

Certification: ijb 2019 0351 [Generate Certificate]

Abstract

Helicobacter pylori (H. pylori) are recognized as fastidious chronic bacterial infections in humans, commonly leads to gastritis, peptic ulcer, gastric cancers and gastric malt lymphoma. Currently, around the globe a massive increase of gastric cancer occurs. It could be reduced by identifying and eradicating of H. Pylori, therefore it is an urgent need to develop an accurate diagnostic method. The current study focused on the diagnosis of H. pylori infection without performing endoscopy highlight the importance of noninvasive tests. In first step molecular studies for H. pylori infection were conducted on genomic DNA that were extracted from salivary secretions to detect H. pylori in saliva by targeting two genes such as 16srRNA and Urec gene but we were failed to detect any H. pylori DNA in saliva. In second step we determined the accuracy of H. pylori culture and then its PCR sequencing. Saliva samples from 60 anti-H. pylori-positive patients were cultured on H. pylori selective media, then colonies were isolated and screened through different biochemical tests. Moreover, H. pylori colonies were amplified on PCR using 16srRNA for validation, our preliminary results were further confirmed on gel electrophoresis and automated sequencer ongoing. Moreover, this study in the future will give information for more accurate and molecular diagnosis of H. pylori infection through noninvasive methods.

| Views 41 |

| Views 41 |

PCR based identification of Helicobacter pylori infection in saliva samples

Abu S, Ruba S, Azmi DH, Dhia SH, Hala IAD. 2014. “Isolation and detection of Helicobacter pylori from patients suffering from peptic ulcer using biochemical tests and molecular techniques.” American Journal of Biochemistry & Biotechnology 58.

Banatvala N, Lopez CR, Robert O, Yasin A, Gareth D, Jeremy H, Roger F. 1993. “Helicobacter pylori in dental plaque. The Lancet 341, 380.

Blanchard TG, John GN. 2012. Laboratory maintenance of Helicobacter species. Current protocols in microbiology 24, 8B-1.

Cheng LH, Webberley M, Evans M, Hanson N, Brown R. 1996. Helicobacter pylori in dental plaque and gastric mucosa. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontics 81, 421-423.

Cutler AF, Havstad S, Ma CK, Blaser MJ, Perez-Perez GI, Schubert TT. 1995. Accuracy of invasive and noninvasive tests to diagnose Helicobacter pylori infection. Gastroenterology             109, 136-141.

Desai HG, Gill HH, Shankaran K, Mehta PR, Prabhu SR. 1991. Dental plaque: a permanent reservoir of Helicobacter pylori? Scandinavian journal of gastroenterology 26, 1205-1208.

Estrada-g IC, Lamb FI, Colston MJ, Robert AC. 1988. Partial nucleotide sequence of 16S ribosomal RNA isolated from armadillo-grown Mycobacterium leprae. Microbiology 134, 1449-1453.

Eusebi, Leonardo H, Rocco MZ, Franco. 2014. Epidemiology of Helicobacter pylori infection. Helicobacter 19, 1-5.

Ferguson DA, Chuanfu LI, Patel NR, Mayberry WR, Chi DS, Thomas E. 1993. Isolation of Helicobacter pylori from saliva. Journal of clinical microbiology 31, 2802-2804.

Graham, David Y. 2015. Helicobacter pylori update: gastric cancer, reliable therapy, and possible benefits. Gastroenterology 148, 719-731.

Hammar M, Tyszkiewicz T, Wadström T, O’toole PW. 1992. Rapid detection of Helicobacter pylori in gastric biopsy material by polymerase chain reaction. Journal of Clinical Microbiology 30, 54-58.

Ho SA, Hoyle JA, Lewis FA, Secker AD, Cross D, Mapstone NP, Dixon MF, Wyatt JI, Tompkins DS, Taylor GR. 1991. Direct polymerase chain reaction test for detection of Helicobacter pylori in humans and animals. Journal of clinical microbiology 29, 2543-2549.

Khasag O, Boldbaatar G, Tegshee T, Duger D, Dashdorj A, Uchida T, Matsuhisa T, Yamaoka Y. 2018. The prevalence of Helicobacter pylori infection and other risk factors among Mongolian dyspeptic patients who have a high incidence and mortality rate of gastric cancer. Gut pathogens 10, 14.

Kim N, Lim SH, Lee KH, You JY, Kim JM, Lee NR, Jung HC, Song IS, Kim CY. 2000. Helicobacter pylori in dental plaque and saliva. The Korean journal of internal medicine 15, 187.

Kolaylı S, Baltas N, Sahin H, Karaoglu S. 2017. Evaluation of anti-Helicobacter pylori activity and urease inhibition by some Turkish authentic honeys. Journal of food sciences and Engineering 7, 67-73.

Krajden S, Fuksa M, Anderson J, Kempston J, Boccia A, Petrea C, Babida C, Karmali MO, Penner JL. 1989. Examination of human stomach biopsies, saliva, and dental plaque for Campylobacter pylori. Journal of clinical microbiology 27, 1397-1398.

Majmudar P, Shah SM, Dhunjibhoy KR, Desai HG. 1990. Isolation of Helicobacter pylori from dental plaques in healthy volunteers. Indian journal of gastroenterology: official journal of the Indian Society of Gastroenterology 9, 271-272.

Mapstone NP, Lynch DA, Lewis FA, Axon AT, Tompkins DS, Dixon MF, Quirke P. 1993. Identification of Helicobacter pylori DNA in the mouths and stomachs of patients with gastritis using PCR. Journal of clinical pathology 46, 540-543.

Okullu SO, Kocagöz T, Tiftikçi A, Yazgan AS, Tozun N. 2017. A Multiplex-Urease PCR Assay for Detection of Helicobacter pylori Infection Directly from Gastric Biopsy Specimens and Comparison of Multiplex-Urease PCR Results with Rapid Urease Test and Histopathology. ClinICAL MicrobiolOGY 6, 2.

Paster BJ, Bruce J, Floyd ED. 1988. Phylogeny of campylobacters, wolinellas, Bacteroides gracilis, and Bacteroides ureolyticus by 16S ribosomal ribonucleic acid sequencing. International Journal of Systematic and Evolutionary Microbiology 38, 56-62.

Patel SK, Chandra BP, Ashok KJ, Anil KG, Gopal N. 2014. Diagnosis of Helicobacter pylori: what should be the gold standard? World journal of gastroenterology 20, 12847.

Rasmussen LT, De Labio RW, Neto AC, Silva LC, Queiroz VF, Smith MD, Payão SL. 2012. Detection of Helicobacter pylori in gastric biopsies, saliva and dental plaques of dyspeptic patients from Marília, São Paulo, Brazil: presence of vacA and cagA genes. Journal of Venomous Animals and Toxins including Tropical Diseases 18, 180-187.

Saez J, Belda S, Santibáñez M, Rodríguez JC, Sola-Vera J, Galiana A, Ruiz-García M, Brotons A, López-GE, Girona E, Sillero C. 2012. Real-time PCR for diagnosing Helicobacter pylori infection in patients with upper gastrointestinal bleeding: comparison with other classical diagnostic methods. Journal of clinical microbiology 1, 3233-7.

Samarbaf-Zadeh AR, Tajbakhsh S, Moosavian SM, Sadeghi-Zadeh M, Azmi M, Hashemi J, Masjedi-Zadeh A. 2006. Application of fluorescent in situ hybridization (FISH) for the detection of Helicobacter pylori. Medical science monitor 1;12(10), CR426-30.

Sarma A, Hazarika BB, Patgiri SJ, Saikia L, Begum S, Hussain ME. 2016. Isolation of Helicobacter Pylori from Gastric Biopsy Specimens and Evaluation of Common Contaminants Associated with H. Pylori Cultures. Int. J. Res. Prof. 2, 161-4.

Savoldi A, Carrara E, Graham DY, Conti M, Tacconelli E. 2018. Prevalence of antibiotic resistance in Helicobacter pylori: a systematic review and meta-analysis in World Health Organization regions. Gastroenterology 1, 1372-82.

Singh V, Mishra S, Rao GR, Jain AK, Dixit VK, Gulati AK, Mahajan D, McClelland M, Nath G. 2008. Evaluation of nested PCR in detection of Helicobacter pylori targeting a highly conserved gene: HSP60. Helicobacter 13, 30-4.

Tiwari SK, Khan AA, Ahmed KS, Ahmed I, Kauser F, Hussain MA, Ali SM, Alvi A, Habeeb A, Abid Z. 2005. Rapid diagnosis of Helicobacter pylori infection in dyspeptic patients using salivary secretion: a non-invasive approach. Singapore medical journal 46, 224.

Tonkic A, Marija T, Philippe L, Francis M. 2012. Epidemiology and Diagnosis of H elicobacter pylori Infection. Helicobacter 17, 1-8.

Toscano, Evelyn P, Fernanda FM, Dutra-Rulli MP, Luiz Otavio MG, Marcela AP, Silva AE. 2018. Epidemiological and Clinical-Pathological Aspects of Helicobacter pylori Infection in Brazilian Children and Adults. Gastroenterology research and practice.

Vianna JS, Ramis IB, Ramos DF, Von Groll A, Silva PE. 2016. Drug resistance in Helicobacter pylori. Arquivos de gastroenterologia 53, 215-23.

Wang YK, Kuo FC, Liu CJ, Wu MC, Shih HY, Wang SS, Wu JY, Kuo CH, Huang YK, Wu DC. 2015. Diagnosis of Helicobacter pylori infection: Current options and developments. World Journal of Gastroenterology 28;21, 11221.

Komal Aman, Bashir Ahmad, Sajid Ali, Saad Ahmad Khan, Md Motiar Rahman, Farhana Jabeen Shah, Muslim Raza, Huang Linfang.
PCR based identification of Helicobacter pylori infection in saliva samples.
Int. J. Biosci. 15(3), 496-504, September 2019.
https://innspub.net/ijb/pcr-based-identification-helicobacter-pylori-infection-saliva-samples/
Copyright © 2019
By Authors and International Network for
Natural Sciences (INNSPUB)
https://innspub.net
brand
innspub logo
english language editing
  • CALL FOR PAPERS
    CALL FOR PAPERS
    Publish Your Article
  • CALL FOR PAPERS
    CALL FOR PAPERS
    Submit Your Article
INNSPUB on FB
Email Update