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

BMC Gastroenterology

Sitang Gong¹  Huimin Xia¹  Zhenwen Zhou¹  Yongmei Zeng¹  Min Yang¹  Peiyu Chen¹  Lixia Li¹  Lanlan Geng¹  Ding-You Li²  Liya Xiong¹  Zhiying Ou¹ 

[1] Department of Gastroenterology, Guangzhou Women and Children’s Medical Center, Guangzhou Medical College, 9 Jinsui Road, Guangzhou, 510623, China;Department of Pediatrics, University of Missouri Kansas City School of Medicine, Division of Gastroenterology, Children’s Mercy Hospital, 2401 Gillham Road, Kansas City, MO, 64108, USA

关键词: Children;    Dental plaques;    Gastric fluids;    Gastric mucosa;    Helicobacter pylori;    Fluorescence quantitative PCR;   

Others  :  858240
DOI  :  10.1186/1471-230X-13-7

 received in 2012-06-21, accepted in 2013-01-11,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

   
2013 Ou et al; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]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.
• [2]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.
• [3]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.
• [4]Frenck RW Jr, 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.
• [5]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.
• [6]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.
• [7]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.
• [8]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.
• [9]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]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.
• [11]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.
• [12]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.
• [13]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]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.
• [15]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.
• [16]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.
• [17]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.
• [18]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.
• [19]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.
• [20]Ashton-Key M, Diss TC, Isaacson PG: Detection of Helicobacter pylori in gastric biopsy and resection specimens. J Clin Pathol 1996, 49:107-111.
• [21]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.
• [22]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.
• [23]Smith SB, Snow AN, Perry RL, Qasem SA: Helicobacter pylori: to stain or not to stain? Am J Clin Pathol 2012, 137:733-738.
• [24]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.
• [25]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.