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Published in: BMC Gastroenterology 1/2013

Open Access 01-12-2013 | Research article

Evaluation of a new fluorescence quantitative PCR test for diagnosing Helicobacter pylori infection in children

Authors: Zhiying Ou, Liya Xiong, Ding-You Li, Lanlan Geng, Lixia Li, Peiyu Chen, Min Yang, Yongmei Zeng, Zhenwen Zhou, Huimin Xia, Sitang Gong

Published in: BMC Gastroenterology | Issue 1/2013

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Abstract

Background

Numerous diagnostic tests are available to detect Helicobactor pylori (H. pylori). There has been no single test available to detect H. pylori infection reliably. We evaluated the accuracy of a new fluorescence quantitative PCR (fqPCR) for H. pylori detection in children.

Methods

Gastric biopsy specimens from 138 children with gastritis were sent for routine histology exam, rapid urease test (RUT) and fqPCR. 13C-urea breath test (13C-UBT) was carried out prior to endoscopic procedure. Gastric fluids and dental plaques were also collected for fqPCR analysis.

Results

38 children (27.5%) were considered positive for H. pylori infection by gold standard (concordant positive results on 2 or more tests). The remaining 100 children (72.5%) were considered negative for H. pylori. Gastric mucosa fqPCR not only detected all 38 H. pylori positive patients but also detected 8 (8%) of the 100 gold standard-negative children or 11 (10.7%) of the 103 routine histology-negative samples. Therefore, gastric mucosa fqPCR identified 46 children (33.3%) with H. pylori infection, significantly higher than gold standard or routine histology (P<0.01). Both gastric fluid and dental plaque fqPCR only detected 32 (23.2%) and 30 (21.7%) children with H. pylori infection respectively and was significantly less sensitive than mucosa fqPCR (P<0.05) but was as sensitive as non-invasive UBT.

Conclusions

Gastric mucosa fqPCR was more sensitive than routine histology, RUT, 13C-UBT alone or in combination to detect H. pylori infection in children with chronic gastritis. Either gastric fluid or dental plaque PCR is as reliable as 13C-UBT for H. pylori detection.
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Literature
1.
go back to reference Mitchell HM, Li YY, Hu PJ, Liu Q, Chen M, Du GG, Wang ZJ, Lee A, Hazell SL: Epidemiology of Helicobacter pylori in southern China: identification of early childhood as the critical period for acquisition. J Infect Dis. 1992, 166: 149-153. 10.1093/infdis/166.1.149.CrossRefPubMed Mitchell HM, Li YY, Hu PJ, Liu Q, Chen M, Du GG, Wang ZJ, Lee A, Hazell SL: Epidemiology of Helicobacter pylori in southern China: identification of early childhood as the critical period for acquisition. J Infect Dis. 1992, 166: 149-153. 10.1093/infdis/166.1.149.CrossRefPubMed
2.
go back to reference Liu Y, Yue H, Li A, Wang J, Jiang B, Zhang Y, Bai Y: An epidemiologic study on the correlation between oral Helicobacter pylori and gastric H. pylori. Curr Microbiol. 2009, 58: 449-453. 10.1007/s00284-008-9341-3.CrossRefPubMed Liu Y, Yue H, Li A, Wang J, Jiang B, Zhang Y, Bai Y: An epidemiologic study on the correlation between oral Helicobacter pylori and gastric H. pylori. Curr Microbiol. 2009, 58: 449-453. 10.1007/s00284-008-9341-3.CrossRefPubMed
3.
go back to reference Guarner J, Kalach N, Elitsur Y, Koletzko S: Helicobacter pylori diagnostic tests in children: review of the literature from 1999 to 2009. Eur J Pediatr. 2010, 169: 15-25. 10.1007/s00431-009-1033-x.CrossRefPubMed Guarner J, Kalach N, Elitsur Y, Koletzko S: Helicobacter pylori diagnostic tests in children: review of the literature from 1999 to 2009. Eur J Pediatr. 2010, 169: 15-25. 10.1007/s00431-009-1033-x.CrossRefPubMed
4.
go back to reference Frenck RW, Fathy HM, Sherif M, Mohran Z, El Mohammedy H, Francis W, Rockabrand D, Mounir BI, Rozmajzl P, Frierson HF: Sensitivity and specificity of various tests for the diagnosis of Helicobacter pylori in Egyptian children. Pediatrics. 2006, 118: e1195-1202. 10.1542/peds.2005-2925.CrossRefPubMed Frenck RW, Fathy HM, Sherif M, Mohran Z, El Mohammedy H, Francis W, Rockabrand D, Mounir BI, Rozmajzl P, Frierson HF: Sensitivity and specificity of various tests for the diagnosis of Helicobacter pylori in Egyptian children. Pediatrics. 2006, 118: e1195-1202. 10.1542/peds.2005-2925.CrossRefPubMed
5.
go back to reference Koletzko S, Jones NL, Goodman KJ, Gold B, Rowland M, Cadranel S, Chong S, Colletti RB, Casswall T, Elitsur Y, Guarner J, Kalach N, Madrazo A, Megraud F, Oderda G: H pylori Working Groups of ESPGHAN and NASPGHAN: evidence-based guidelines from ESPGHAN and NASPGHAN for Helicobacter pylori infection in children. J Pediatr Gastroenterol Nutr. 2011, 53: 230-243.PubMed Koletzko S, Jones NL, Goodman KJ, Gold B, Rowland M, Cadranel S, Chong S, Colletti RB, Casswall T, Elitsur Y, Guarner J, Kalach N, Madrazo A, Megraud F, Oderda G: H pylori Working Groups of ESPGHAN and NASPGHAN: evidence-based guidelines from ESPGHAN and NASPGHAN for Helicobacter pylori infection in children. J Pediatr Gastroenterol Nutr. 2011, 53: 230-243.PubMed
6.
go back to reference Gisbert JP, Abraira V: Accuracy of Helicobacter pylori diagnostic tests in patients with bleeding peptic ulcer: a systematic review and meta-analysis. Am J Gastroenterol. 2006, 101: 848-863. 10.1111/j.1572-0241.2006.00528.x.CrossRefPubMed Gisbert JP, Abraira V: Accuracy of Helicobacter pylori diagnostic tests in patients with bleeding peptic ulcer: a systematic review and meta-analysis. Am J Gastroenterol. 2006, 101: 848-863. 10.1111/j.1572-0241.2006.00528.x.CrossRefPubMed
7.
go back to reference Yoo JY, Kim N, Park YS, Hwang JH, Kim JW, Jeong SH, Lee HS, Choe C, Lee DH, Jung HC, Song IS: Detection rate of Helicobacter pylori against a background of atrophic gastritis and/or intestinal metaplasia. J Clin Gastroenterol. 2007, 41: 751-755. 10.1097/MCG.0b013e31802c347d.CrossRefPubMed Yoo JY, Kim N, Park YS, Hwang JH, Kim JW, Jeong SH, Lee HS, Choe C, Lee DH, Jung HC, Song IS: Detection rate of Helicobacter pylori against a background of atrophic gastritis and/or intestinal metaplasia. J Clin Gastroenterol. 2007, 41: 751-755. 10.1097/MCG.0b013e31802c347d.CrossRefPubMed
8.
go back to reference Leal YA, Flores LL, Fuentes-Pananá EM, Cedillo-Rivera R, Torres J: 13C-urea breath test for the diagnosis of Helicobacter pylori infection in children: a systematic review and meta-analysis. Helicobacter. 2011, 16: 327-337. 10.1111/j.1523-5378.2011.00863.x.CrossRefPubMed Leal YA, Flores LL, Fuentes-Pananá EM, Cedillo-Rivera R, Torres J: 13C-urea breath test for the diagnosis of Helicobacter pylori infection in children: a systematic review and meta-analysis. Helicobacter. 2011, 16: 327-337. 10.1111/j.1523-5378.2011.00863.x.CrossRefPubMed
9.
go back to reference Choi J, Kim CH, Kim D, Chung SJ, Song JH, Kang JM, Yang JI, Park MJ, Kim YS, Yim JY, Lim SH, Kim JS, Jung HC, Song IS: Prospective evaluation of a new stool antigen test for the detection of Helicobacter pylori, in comparison with histology, rapid urease test, (13)C-urea breath test, and serology. J Gastroenterol Hepatol. 2011, 26: 1053-1059. 10.1111/j.1440-1746.2011.06705.x.CrossRefPubMed Choi J, Kim CH, Kim D, Chung SJ, Song JH, Kang JM, Yang JI, Park MJ, Kim YS, Yim JY, Lim SH, Kim JS, Jung HC, Song IS: Prospective evaluation of a new stool antigen test for the detection of Helicobacter pylori, in comparison with histology, rapid urease test, (13)C-urea breath test, and serology. J Gastroenterol Hepatol. 2011, 26: 1053-1059. 10.1111/j.1440-1746.2011.06705.x.CrossRefPubMed
10.
go back to reference Tiveljung A, Borch K, Jonasson J, Mårdh S, Petersson F, Monstein HJ: Identification of Helicobacter in gastric biopsies by PCR based on 16S rDNA sequences: a matter of little significance for the prediction of H. pylori-associated gastritis?. J Med Microbiol. 1998, 47: 695-704. 10.1099/00222615-47-8-695.CrossRefPubMed Tiveljung A, Borch K, Jonasson J, Mårdh S, Petersson F, Monstein HJ: Identification of Helicobacter in gastric biopsies by PCR based on 16S rDNA sequences: a matter of little significance for the prediction of H. pylori-associated gastritis?. J Med Microbiol. 1998, 47: 695-704. 10.1099/00222615-47-8-695.CrossRefPubMed
11.
go back to reference Zsikla V, Hailemariam S, Baumann M, Mund MT, Schaub N, Meier R, Cathomas G: Increased rate of Helicobacter pylori infection detected by PCR in biopsies with chronic gastritis. Am J Surg Pathol. 2006, 30: 242-248. 10.1097/01.pas.0000186397.18206.1b.CrossRefPubMed Zsikla V, Hailemariam S, Baumann M, Mund MT, Schaub N, Meier R, Cathomas G: Increased rate of Helicobacter pylori infection detected by PCR in biopsies with chronic gastritis. Am J Surg Pathol. 2006, 30: 242-248. 10.1097/01.pas.0000186397.18206.1b.CrossRefPubMed
12.
go back to reference Molnar B, Szoke D, Ruzsovics A, Tulassay Z: Significantly elevated Helicobacter pylori density and different genotype distribution in erosions as compared with normal gastric biopsy specimen detected by quantitative real-time PCR. Eur J Gastroenterol Hepatol. 2008, 20: 305-313. 10.1097/MEG.0b013e3282f2fda4.CrossRefPubMed Molnar B, Szoke D, Ruzsovics A, Tulassay Z: Significantly elevated Helicobacter pylori density and different genotype distribution in erosions as compared with normal gastric biopsy specimen detected by quantitative real-time PCR. Eur J Gastroenterol Hepatol. 2008, 20: 305-313. 10.1097/MEG.0b013e3282f2fda4.CrossRefPubMed
13.
go back to reference Belda S, Saez J, Santibáñez M, Rodríguez JC, Galiana A, Sola-Vera J, Ruiz-García M, Brotons A, López-Girona E, Girona E, Sillero C, Royo G: Quantification of Helicobacter pylori in gastric mucosa by real-time polymerase chain reaction: comparison with traditional diagnostic methods. Diagn Microbiol Infect Dis. 2012, Epub ahead of print Belda S, Saez J, Santibáñez M, Rodríguez JC, Galiana A, Sola-Vera J, Ruiz-García M, Brotons A, López-Girona E, Girona E, Sillero C, Royo G: Quantification of Helicobacter pylori in gastric mucosa by real-time polymerase chain reaction: comparison with traditional diagnostic methods. Diagn Microbiol Infect Dis. 2012, Epub ahead of print
14.
go back to reference Kindermann A, Demmelmair H, Koletzko B, Krauss-Etschmann S, Wiebecke B, Koletzko S: Influence of age on 13C-urea breath test results in children. J Pediatr Gastroenterol Nutr. 2000, 30: 85-91. 10.1097/00005176-200001000-00023.CrossRefPubMed Kindermann A, Demmelmair H, Koletzko B, Krauss-Etschmann S, Wiebecke B, Koletzko S: Influence of age on 13C-urea breath test results in children. J Pediatr Gastroenterol Nutr. 2000, 30: 85-91. 10.1097/00005176-200001000-00023.CrossRefPubMed
15.
go back to reference Mana F, Franken PR, Ham HR, Urbain D: Cut-off point, timing and pitfalls of the 13C-urea breath test as measured by infrared spectrometry. Dig Liver Dis. 2001, 33: 30-35. 10.1016/S1590-8658(01)80132-8.CrossRefPubMed Mana F, Franken PR, Ham HR, Urbain D: Cut-off point, timing and pitfalls of the 13C-urea breath test as measured by infrared spectrometry. Dig Liver Dis. 2001, 33: 30-35. 10.1016/S1590-8658(01)80132-8.CrossRefPubMed
16.
go back to reference Kim GA, Lee MS, Sun Y, Lee BD, Lee JI, Lee JH, Kwon ST: Characterization of cold-active uracil-DNA glycosylase from Bacillus sp. HJ171 and its use for contamination control in PCR. Appl Microbiol Biotechnol. 2008, 80: 785-794. 10.1007/s00253-008-1585-0.CrossRefPubMed Kim GA, Lee MS, Sun Y, Lee BD, Lee JI, Lee JH, Kwon ST: Characterization of cold-active uracil-DNA glycosylase from Bacillus sp. HJ171 and its use for contamination control in PCR. Appl Microbiol Biotechnol. 2008, 80: 785-794. 10.1007/s00253-008-1585-0.CrossRefPubMed
17.
go back to reference Lage AP, Godfroid E, Fauconnier A, Burette A, Butzler JP, Bollen A, Glupczynski Y: Diagnosis of Helicobacter pylori infection by PCR: comparison with other invasive techniques and detection of cagA gene in gastric biopsy specimens. J Clin Microbiol. 1995, 33: 2752-2756.PubMedPubMedCentral Lage AP, Godfroid E, Fauconnier A, Burette A, Butzler JP, Bollen A, Glupczynski Y: Diagnosis of Helicobacter pylori infection by PCR: comparison with other invasive techniques and detection of cagA gene in gastric biopsy specimens. J Clin Microbiol. 1995, 33: 2752-2756.PubMedPubMedCentral
18.
go back to reference Ramírez-Lázaro MJ, Lario S, Casalots A, Sanfeliu E, Boix L, García-Iglesias P, Sánchez-Delgado J, Montserrat A, Bella-Cueto MR, Gallach M, Sanfeliu I, Segura F, Calvet X: Real-time PCR improves Helicobacter pylori detection in patients with peptic ulcer bleeding. PLoS One. 2011, 6: e20009-10.1371/journal.pone.0020009.CrossRefPubMedPubMedCentral Ramírez-Lázaro MJ, Lario S, Casalots A, Sanfeliu E, Boix L, García-Iglesias P, Sánchez-Delgado J, Montserrat A, Bella-Cueto MR, Gallach M, Sanfeliu I, Segura F, Calvet X: Real-time PCR improves Helicobacter pylori detection in patients with peptic ulcer bleeding. PLoS One. 2011, 6: e20009-10.1371/journal.pone.0020009.CrossRefPubMedPubMedCentral
19.
go back to reference Weiss J, Tsang TK, Meng X, Zhang H, Kilner E, Wang E, Watkin W: Detection of Helicobacter pylori gastritis by PCR: correlation with inflammation scores and immunohistochemical and CLOtest findings. Am J Clin Pathol. 2008, 129: 89-96. 10.1309/APMPEP54G7PN958G.CrossRefPubMed Weiss J, Tsang TK, Meng X, Zhang H, Kilner E, Wang E, Watkin W: Detection of Helicobacter pylori gastritis by PCR: correlation with inflammation scores and immunohistochemical and CLOtest findings. Am J Clin Pathol. 2008, 129: 89-96. 10.1309/APMPEP54G7PN958G.CrossRefPubMed
20.
go back to reference Ashton-Key M, Diss TC, Isaacson PG: Detection of Helicobacter pylori in gastric biopsy and resection specimens. J Clin Pathol. 1996, 49: 107-111. 10.1136/jcp.49.2.107.CrossRefPubMedPubMedCentral Ashton-Key M, Diss TC, Isaacson PG: Detection of Helicobacter pylori in gastric biopsy and resection specimens. J Clin Pathol. 1996, 49: 107-111. 10.1136/jcp.49.2.107.CrossRefPubMedPubMedCentral
21.
go back to reference Jonkers D, Stobberingh E, de Bruine A, Arends JW, Stockbrügger R: Evaluation of immunohistochemistry for the detection of Helicobacter pylori in gastric mucosal biopsies. J Infect. 1997, 35: 149-514.CrossRefPubMed Jonkers D, Stobberingh E, de Bruine A, Arends JW, Stockbrügger R: Evaluation of immunohistochemistry for the detection of Helicobacter pylori in gastric mucosal biopsies. J Infect. 1997, 35: 149-514.CrossRefPubMed
22.
go back to reference Orhan D, Kalel G, Saltik-Temizel IN, Demir H, Bulun A, Karaağaoğlu E, Cağlar M: Immunohistochemical detection of Helicobacter pylori infection in gastric biopsies of urea breath test-positive and -negative pediatric patients. Turk J Pediatr. 2008, 50: 34-39.PubMed Orhan D, Kalel G, Saltik-Temizel IN, Demir H, Bulun A, Karaağaoğlu E, Cağlar M: Immunohistochemical detection of Helicobacter pylori infection in gastric biopsies of urea breath test-positive and -negative pediatric patients. Turk J Pediatr. 2008, 50: 34-39.PubMed
23.
go back to reference Smith SB, Snow AN, Perry RL, Qasem SA: Helicobacter pylori: to stain or not to stain?. Am J Clin Pathol. 2012, 137: 733-738. 10.1309/AJCP8DGTAVG7MBMT.CrossRefPubMed Smith SB, Snow AN, Perry RL, Qasem SA: Helicobacter pylori: to stain or not to stain?. Am J Clin Pathol. 2012, 137: 733-738. 10.1309/AJCP8DGTAVG7MBMT.CrossRefPubMed
24.
go back to reference Komen NA, Bertleff MJ, van Doorn LJ, Lange JF, de Graaf PW: Helicobacter genotyping and detection in peroperative lavage fluid in patients with perforated peptic ulcer. J Gastrointest Surg. 2008, 12: 555-560. 10.1007/s11605-007-0303-z.CrossRefPubMed Komen NA, Bertleff MJ, van Doorn LJ, Lange JF, de Graaf PW: Helicobacter genotyping and detection in peroperative lavage fluid in patients with perforated peptic ulcer. J Gastrointest Surg. 2008, 12: 555-560. 10.1007/s11605-007-0303-z.CrossRefPubMed
25.
go back to reference Tsami A, Petropoulou P, Kafritsa Y, Mentis YA, Roma-Giannikou E: The presence of Helicobacter pylori in dental plaque of children and their parents: is it related to their periodontal status and oral hygiene?. Eur J Paediatr Dent. 2011, 12: 225-230.PubMed Tsami A, Petropoulou P, Kafritsa Y, Mentis YA, Roma-Giannikou E: The presence of Helicobacter pylori in dental plaque of children and their parents: is it related to their periodontal status and oral hygiene?. Eur J Paediatr Dent. 2011, 12: 225-230.PubMed
Metadata
Title
Evaluation of a new fluorescence quantitative PCR test for diagnosing Helicobacter pylori infection in children
Authors
Zhiying Ou
Liya Xiong
Ding-You Li
Lanlan Geng
Lixia Li
Peiyu Chen
Min Yang
Yongmei Zeng
Zhenwen Zhou
Huimin Xia
Sitang Gong
Publication date
01-12-2013
Publisher
BioMed Central
Published in
BMC Gastroenterology / Issue 1/2013
Electronic ISSN: 1471-230X
DOI
https://doi.org/10.1186/1471-230X-13-7

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