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ISSN: 1747-0862
Journal of Molecular and Genetic Medicine
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Diagnostic Accuracy of PCR-Based Detection Tests for Helicobacter Pylori in Otitis Media: A Meta-Analysis

Fakher Rahim*

PhD in Molecular Medicine, Toxicology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

Corresponding Author:
Fakher Rahim
Toxicology Research Center
Ahvaz Jundishapur University of Medical sciences, Ahvaz, Iran
Tel: +986113367562
E-mail: Bioinfo2003@ajums.ac.ir/F-rahim@Razi.Tums.ac.ir

Received date: June 20, 2013; Accepted date: August 05, 2013; Published date: August 07, 2013

Citation: Rahim F (2013) Diagnostic Accuracy of PCR-Based Detection Tests for Helicobacter Pylori in Otitis Media: A Meta-Analysis. J Mol Genet Med 7:62. doi:10.4172/1747-0862.1000062

Copyright: © 2013 Rahim F. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Keywords

Diagnostic accuracy; Meta-analysis; H. Pylori infection; Otitis media with effusion (OME); Polymerase chain reaction (PCR); Histology

Introduction

Helicobacter pylori (H. Pylori) infection has serious consequences such as peptic ulcers and gastric cancer [1-3]. Histologic identification remains the gold standard in diagnosing H. pylori infection [4]. The urea breath test, polymerase chain reaction (PCR), and CLO are also the most reliable diagnostic techniques for H. pylori infection [5]. Some researchers reported the association between H. pylori infection and upper respiratory diseases, including chronic rhino sinusitis, chronic otitis media, and chronic otitis media with effusion [6-12], but little is known about the true colonization and the localization of these bacteria in the upper respiratory tract tissue.

Otitis media with effusion (OME) is a more common condition in persons with poor Eustachian tube function, in which the pathogenesis still remains unknown. Viral or bacterial infections, autoimmunity, allergy, gastro esophageal reflux possibly play a role in the pathogenesis of OME [13]. Recently, the heritability and genetic determinants of OME were also studied [14,15]. Epidemiological evidences have suggested the possible relationship of H. Pylori with the OME [16-18]. Over the last decade, several molecular techniques have been developed for targeting various microbial genes [19,20]. Polymerase chain reaction (PCR) is one of the most common molecular techniques used in detecting H. Pylori infection [21-23].To the best of our knowledge, the diagnostic accuracy of PCR-based methods for OME-infected patients has not been systematically reviewed and synthesized. Thus, we conducted a meta-analysis and systematic review to summarize the evidence on diagnostic accuracy of PCR-based tests compared to the histological methods in the patients with OME.

Materials and Methods

Literature review

We performed a systematic search without a language limitation in the Medline, Scopus and ISI web of science, covering all published papers up to March 2013, with the following combination of Mesh standardized keywords: (("Helicobacter pylori"[Mesh]) AND (“Otitis Media"[Mesh] OR "Otitis Media with Effusion"[Mesh] OR "Otitis Media, Suppurative"[Mesh])) OR "Otolaryngology"[Mesh]. We reviewed potentially selected publications through titles and abstract screening and then collected the most related publications for a closer evaluation. Besides, the reference lists of the selected papers were also screened for other potential articles that possibly have been missed in the initial search.

Inclusion and exclusion criteria

The PCR techniques were compared to the histological tests as the gold standard in diagnosing the H. pylori infection. Those studies, which concern the association of H. pylori infection with OME risk using PCR and histological methods, offer the size of the sample, and the information that can help infer the results, were included. Accordingly, the reviews and duplicated publications were excluded.

Data extraction

Full papers of any titles/abstracts that appeared to be relevant were obtained where possible and the relevance of each study independently was assessed according to the aforementioned inclusion and exclusion criteria.

Assessment of study quality

We measured the quality of studies using the Quality Assessment of Studies of Diagnostic Accuracy Approach-QUADAS [24], a 14-item specifically developed tool, to judge the quality of primary studies.

Statistical analysis

The diagnostic odds ratio (DOR) and symmetric summary receiver operating characteristic (sROC) of OME-associated H. pylori infection, were estimated for each study. The DOR and its 95% confidence interval (CI) to each study was plotted against the number of participants for detecting any possible sample size biases. The heterogeneity was tested based on the I-squared values, with values less than 25%, 25% to 50%, and greater than 50% indicating low, moderate and high heterogeneity, respectively [25]. Using the bivariate method according to Reitsma et al. [26], pooled sensitivity, specificity and 95% confidence intervals were estimated for each diagnostic test. Publication biases were observed, using the relationship between the diagnostic odds ratio (DOR) and the effective sample size (ESS) [27]. Statistical analysis was undertaken using the program STATA 11.0 software. The Meta-Disk was used to calculate individual and pooled diagnostic OR, sensitivity, specificity, negative likelihood ratio, positive likelihood ratio [28]. P <0.05 was considered statistically significant.

Results

From the literature searches, we included eight relevant studies compromising 259 case of OME, which study selection flow is shown in figure 1.Only two studies were stated sensitivity and specificity. Table 1 summarizes the characteristics of eight studies. The quality analysis using QUADAS tool showed that six out of eight studies (75%) fulfilled more than seven criteria. The individual and combined sensitivity and specificity estimations for the PCR test, including all selected studies were assessed (Figure 2). Overall, sensitivity and specificity varied among studies of a given PCR test. The pooled sensitivity and specificity of PCR test in diagnosing H. Pylori infection in patient with OME were 71% (95%CI: 61%-80%, I2: 0.0%) and 81% (95%CI: 76% - 86%,I2: 59.9%), respectively. Pooled positive likelihood ratio (PLR) and negative likelihood ratio (NLR) for PCR were 3.61(95%CI: 2.34 – 5.59, I2: 44.5%) and 0.42 (95%CI: 0.31 – 0.57, I2: 0.1%), respectively (Figure 3).The sROC plots of eight selected studies and estimated DOR (95% confidence interval) are shown in figure 4A. For DOR analysis, the pooled accuracy of PCR was 10.78 (95%CI: 5.95 – 19.53, I2: 0.0%) in diagnosing H. Pylori infection in patient with OME (Figure 4B). Egger’s test 2-sided p-value was larger than 0.05, suggesting absence of publication bias for all tests.

molecular-genetic-medicine-included-excluded-studies

Figure 1: The flow diagram of included/excluded studies.

molecular-genetic-medicine-random-effects-model

Figure 2: Estimates of sensitivity (Top) and specificity (Bottom) (95% confidence interval) of PCR. Combined results are shown using bothoptions: fixed and random effects model. When both results are similar with low heterogeneity, both can be used.

molecular-genetic-medicine-positive-likelihood-ratio

Figure 3: Estimates of positive likelihood ratio (PLR, Top) and negativelikelihood ratio (NLR, Bottom) (95% confidence interval) of PCR. Combined results are shown using both options: fixed and random effects model. When both results are similar with low heterogeneity, both can be used.

molecular-genetic-medicine-combined-studies

Figure 4: A, sROC curve for PCR in patient with OME; B, DOR of individual and combined studies.

molecular-genetic-medicine-meta-analysis

PCR, Polymerase Chain Reaction
Table 1: H. pylori infection status among OME cases included in the meta-analysis.

Discussion

The H. Pylori infection diagnosis usually relies on the serology and histologic identification in the past [4]. However, the serology and histologic identification are not fast and reliable enough [29-33].The PCR-based molecular diagnostic techniques have been used for testing H. Pylori infections for about a decade [34,35]. Our meta-analysis revealed some concerns using the available evidences, which basically reflects the experience with PCR-based molecular diagnosis of H. Pylori infection among patient with OME, including the limitation of available evidences and a considerable variability among the available studies. Furthermore, the accuracy of molecular methods such as PCR is great enough, although specificity was generally high, and there is unpredictably variation in specificity among studies evaluating the same test.

Several limitations have been introduced for PCR-based molecular diagnostic, including presence false-negative and false-positive results due to the PCR inhibitors in the samples and easy contamination, and a difficulty in obtaining the worthy samples. Diagnostic accuracy of PCR may decrease at seven days after onset of the disease compared to the histologic identification and serology [36,37].

This meta-analysis addressed a moderate heterogeneity that likely was due to the differences between studies in the design (test protocol, definition of a positive result), geographical location (diverse H. Pylori infection prevalence, local strain dominance) and ethnicity (Asian and American).

This meta-analysis also showed that PCR-based molecular diagnostic method may generate consistent result with high and more variable specificity than sensitivity. Possible descriptions for these differences may include the standard control of PCR and threshold, types of the subjects and time point for sampling. Because of the absence of relative limited high quality studies and small number of published papers, this systematic review and meta-analysis have limitations, which need to be cited with cautious. Furthermore, because of only eight studies included in this review, this meta-analysis could not give strong enough evidence; thus, the findings should to be addressed in further and additional research.

Conclusion

This review showed statistically significant differences in the diagnostic accuracy between the PCR and histological tests. This metaanalysis also suggests a higher sensitivity and specificity of PCR-based molecular diagnostic method for H. Pylori infection in OME patients compared to the histological tests.

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