**Pathology and Repair**

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[1] Nanci A. Ten Cate's Oral Histology Development, Structure and Function. 8th ed.

[2] Kumar GS. Orban's Oral Histology and Embryology. 13th ed. Elsevier; 2014. pp. 291-312 [3] Eroschenko VP. DiFiore's Atlas of Histology with Functional Correlations. 11th ed.

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[5] Cutler LS, Gremski W. Epithelial-mesenchymal interaction in the development of the

[7] Nedvetsky PI, Emmerson E, Finley JK, Ettinger A, Cruz-Pacheo N, et al. Parasympathetic innervation regulates tubulogenesis in the developing salivary gland. Developmental

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[10] Dodds MW, DA J, Yeh CK. Health benefits of saliva: A review. Journal of Dentistry.

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Neck and Brain. 4th ed. Satish Kumar Jain; 2006. pp. 133-138; 157-162

salivary gland. Critical Reviews in Oral Biology and Medicine. 1991;**2**(1):1

**Chapter 5**

**Provisional chapter**

**A Study of the Correlation between Bacterial Culture**

**A Study of the Correlation between Bacterial Culture** 

*Helicobacter pylori* (*H. pylori*) is one of the most common chronic bacterial infections in the world, and it is currently estimated that approximately half of the world's population is infected with the bacterium. The correct diagnosis and effective treatment of *H. pylori* gastric infection are essential in controlling this condition. The available diagnostic methods have advantages and limitations related to factors such as age of patients, technical difficulty level, costs and extensive accessibility in hospitals. The eradication therapy of *H. pylori* infection is still a challenge for gastroenterologists. One of the main causes of failure in *H. pylori* eradication is antibiotic resistance. Biopsy cultures are the most widely used methods among the antimicrobial susceptibility tests. In case of a negative culture, *H. pylori* can be clearly recognised in histological sections. The sensitivity and specificity of histology for the diagnosis depend on clinical settings, density of colonisation and the experience of the histopathologist. A prospective study was performed in order to analyse patients with *H. pylori* gastric infection with positive histology and positive culture versus positive histology and negative culture.

> © 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution,

© 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,

distribution, and reproduction in any medium, provided the original work is properly cited.

and reproduction in any medium, provided the original work is properly cited.

*Helicobacter pylori* (*H. pylori*) is a Gram-negative spiral-shaped bacterium which colonises the gastrointestinal tract of nearly half of the world's population, causing local inflammation in

DOI: 10.5772/intechopen.80257

**and Histological Examination in Children with**

**and Histological Examination in Children with** 

*Helicobacter pylori* **Gastritis**

*Helicobacter pylori* **Gastritis**

Mihaela Bălgrădean

**Abstract**

**1. Introduction**

Mihaela Bălgrădean

Felicia Galoș, Gabriela Năstase, Cătălin Boboc, Cristina Coldea, Mălina Anghel, Anca Orzan and

Felicia Galoș, Gabriela Năstase, Cătălin Boboc, Cristina Coldea, Mălina Anghel, Anca Orzan and

Additional information is available at the end of the chapter

**Keywords:** *Helicobacter pylori* culture, histology, children

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.80257

#### **A Study of the Correlation between Bacterial Culture and Histological Examination in Children with** *Helicobacter pylori* **Gastritis A Study of the Correlation between Bacterial Culture and Histological Examination in Children with**  *Helicobacter pylori* **Gastritis**

DOI: 10.5772/intechopen.80257

Felicia Galoș, Gabriela Năstase, Cătălin Boboc, Cristina Coldea, Mălina Anghel, Anca Orzan and Mihaela Bălgrădean Felicia Galoș, Gabriela Năstase, Cătălin Boboc, Cristina Coldea, Mălina Anghel, Anca Orzan and Mihaela Bălgrădean

Additional information is available at the end of the chapter Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.80257

#### **Abstract**

*Helicobacter pylori* (*H. pylori*) is one of the most common chronic bacterial infections in the world, and it is currently estimated that approximately half of the world's population is infected with the bacterium. The correct diagnosis and effective treatment of *H. pylori* gastric infection are essential in controlling this condition. The available diagnostic methods have advantages and limitations related to factors such as age of patients, technical difficulty level, costs and extensive accessibility in hospitals. The eradication therapy of *H. pylori* infection is still a challenge for gastroenterologists. One of the main causes of failure in *H. pylori* eradication is antibiotic resistance. Biopsy cultures are the most widely used methods among the antimicrobial susceptibility tests. In case of a negative culture, *H. pylori* can be clearly recognised in histological sections. The sensitivity and specificity of histology for the diagnosis depend on clinical settings, density of colonisation and the experience of the histopathologist. A prospective study was performed in order to analyse patients with *H. pylori* gastric infection with positive histology and positive culture versus positive histology and negative culture.

**Keywords:** *Helicobacter pylori* culture, histology, children

## **1. Introduction**

*Helicobacter pylori* (*H. pylori*) is a Gram-negative spiral-shaped bacterium which colonises the gastrointestinal tract of nearly half of the world's population, causing local inflammation in

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

the stomach and duodenum and inducing a systemic humoral immune response [1]. *H. pylori* survival in the acidic gastric environment is mediated by mechanisms such as activity of the urease enzyme, which catalyses and hydrolyses urea to form carbon dioxide and ammonia, producing a neutral environment that is essential for its survival. The primary routes of transmission are considered to be faecal-oral and oral-oral, but some indirect evidences report that the infection can also be acquired by drinking water and by other environmental sources [2, 3].

before the beginning of the study, previous intestinal surgery (except for polypectomy and appendectomy), concomitant severe disease (heart, lungs, kidney and endocrine diseases)

A Study of the Correlation between Bacterial Culture and Histological Examination in Children…

http://dx.doi.org/10.5772/intechopen.80257

81

All patients underwent endoscopy with biopsy specimens for histology (one for the antrum, one for the corpus). One sample from the antrum was used for rapid urease test. Two additional biopsies were taken from the antrum for bacterial culture. The samples were placed into separate vials, previously identified, containing the appropriate medium for each test.

This procedure was performed in patients with a minimum of 10 hours of fasting, under general anaesthesia or conscious sedation. Vital signs were continuously monitored for the

Written informed consent was obtained from the parent or guardian of each child included

The biopsy specimens collected for bacterial culture were transported in commercial selective transport *H. pylori* medium, Portagerm pylori (BioMérieux SA, Marcy l'Etoile, France), and were inoculated after a few hours onto selective medium pylori agar (BioMérieux Italia). The plates were incubated under microaerobic condition at 37° for 72 hours. Once incubated, the colonies resembling *H. pylori* were identified by Gram stain and by oxidase, catalase and urease tests. Suspensions from the primary plates were prepared in sterile solution to perform an E-test on pylori agar. An agar plate was streaked in three directions with a swab dipped into each bacterial suspension to produce a lawn of growth; an E-test strip (E-test; AB Biodisk, Solna, Sweden) was placed each onto separate plates, which was immediately incubated in a microaerobic atmosphere at 37° for 72 hours. Isolated strains were tested for amoxicillin, clarithromycin, metronidazole and levofloxacin resistance following the recommendations of

A biopsy of the gastric body and antrum was fixed in a solution of formaldehyde 10%. Subsequently, the gastric mucosa samples were processed, following the usual steps of dehy-

Two stains were used for histological study: haematoxylin-eosin and Giemsa. Haematoxylin-eosin stain was used to evaluate inflammatory cells and *H. pylori* (**Figure 1**). Giemsa stain was needed when haematoxylin-eosin stain failed to identify the bacterium (**Figure 2**). Giemsa stain is the preferred stain for detecting *H. pylori* because of its technical simplicity, high sensitivity and low cost.

the European Committee on Antimicrobial Susceptibility Testing.

and smoking or alcohol consumption among adolescents.

The study was approved by the ethics committee.

The first sample was used for bacterial culture.

**2.2. Endoscopy**

entire procedure.

**2.3. Bacterial culture**

in the study.

**2.4. Histology**

dration and paraffin embedding.

*H. pylori* represents a key factor in the aetiology of various gastrointestinal diseases, ranging from asymptomatic chronic active gastritis to peptic ulceration, gastric adenocarcinoma and gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Other diseases caused by the pathogen are iron deficiency anaemia, chronic idiopathic thrombocytopenic purpura and growth retardation. *H. pylori* was classified as a class I human carcinogen by the World Health Organization in 1994.

Numerous diagnostic tests are available for detecting *H. pylori* infection: invasive techniques, which means endoscopy with biopsies for a rapid urease test (RUT), histology and culture and noninvasive techniques, such as serology, 13C-Urea breath test (13C-UBT) and stool antigen test. There is no single method to detect *H. pylori* infection reliably and accurately. The choice of the diagnostic method depends on patients' age and complaints, technical difficulty level, costs and extensive accessibility in hospitals.

Two tests are recommended to define *H. pylori* status in children: bacterial culture of gastric biopsy and histology [4]. Bacterial culture of gastric biopsy has 100% specificity, but its sensitivity is low. *H. pylori* can be cultured from gastric biopsies, although this method often presents some difficulties. *H. pylori* soon loses viability when exposed to the environment, and biopsies should be cultured quickly. If it is not possible, a transport media may be used. Histology provides an excellent diagnostic accuracy, allowing for the detection of the bacteria as well as for the grading of gastritis. The sensitivity and specificity of histology for the diagnosis depend on clinical settings, density of colonisation and experience of the histopathologist [5].

The aims of our study were to assess the histological findings and to compare them with the results of bacterial cultures, obtained through gastric biopsy, in children with *H. pylori* gastritis. We also wanted to find out the possible factors that may influence bacterial culture outcomes.
