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Minerva Biotecnologica 2019 December;31(4):116-21

DOI: 10.23736/S1120-4826.19.02558-8


lingua: Inglese

Comparison of polymerase chain reaction-based genotyping of Helicobacter pylori by direct polymerase chain reaction from biopsies and cultures from patients with dyspepsia in Nigeria

Stella I. SMITH 1 , Tolu JOLAIYA 2, Muinah FOWORA 1, Rose UGIAGBE 3, Charles ONYEKWERE 4, Ifeanyi AGBO 3, Olufunmilayo LESI 5, Pia PALAMIDES 6, Abimbola ADEDEJI 7, Dennis NDUBUBA 8, Olusegun ADEKANLE 8, Isaac ADELEYE 2, Moses BAMIDELE 1, Favour NGOKA 1, Idowu AYODEJI 9, Henry NJOM 9, Rinaldo PELLICANO 10

1 Department of Molecular Biology and Biotechnology, Nigerian Institute of Medical Research, Yaba, Nigeria; 2 Department of Microbiology, University of Lagos, Akoka, Nigeria; 3 Department of Medicine, University of Benin Teaching Hospital, Benin, Nigeria; 4 Department of Medicine, Lagos State University Teaching Hospital, Ikeja, Nigeria; 5 Department of Medicine, Lagos University Teaching Hospital, Idi-Araba, Nigeria; 6 Medical Microbiology and Hospital Epidemiology, Faculty of Medicine, Max von Pettenkofer Institute, LMU Munich, Munich, Germany; 7 Unit of Monitoring and Evaluation, Nigerian Institute of Medical Research, Yaba, Nigeria; 8 Department of Medicine, Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Nigeria; 9 Department of Biochemistry and Microbiology, Faculty of Science and Agriculture, University of Fort Hare, Eastern Cape, South Africa; 10 Unit of Gastroenterology, Molinette-SGAS Hospital, Turin, Italy

BACKGROUND: Helicobacter pylori infects 50% of humans worldwide. Since this bacterium is the main cause of gastritis and peptic ulcer disease, its accurate diagnosis and eradication are crucial steps in the management of these diseases. When endoscopy is required, there is an increasing interest in the real time diagnosis of H. pylori infection and its immediate treatment. Aim of this study was to determine the most accurate method for genotyping H. pylori in Nigeria, comparing results obtained from analysis of bacterial isolates versus those obtained from direct analysis on gastric biopsies.
METHODS: Bioptic specimens, obtained during upper gastrointestinal endoscopy from 125 dyspeptic subjects, after informed consent, were analyzed to detect virulence genotyping of H. pylori, directly or on isolates obtained by culture.
RESULTS: Out of 125 patients, 100 (80%) were positive for H. pylori by direct polymerase chain reaction (PCR) from bioptic specimens while culture was positive in 47 (37.6%). Results of amplified genes showed that comparing 47 samples, positive by culture and direct PCR, the agreement ranged from fair to good. In terms of age group and virulence genotypes, the age group 30-40 years was more common positive for cagA, vacA-s1b, vacA-m1 but not for dupA. However, this result was not statistically significant when compared to the other age-groups.
CONCLUSIONS: The study of the virulence genotypes of H. pylori, using the two methods, direct PCR and PCR after culture, showed that the former was slightly superior to the latter. However, culture remains important for the treatment of patients after failure of antibiotic therapy.

KEY WORDS: Biopsy; Helicobacter pylori; Polymerase chain reaction

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