Effect of Pretreatment With Lactobacillus Gasseri OLL2716 on First-Line Helicobacter Pylori Eradication Therapy

Ryuzo Deguchi; Hidemasa Nakaminami; Emiko Rimbara; Norihisa Noguchi; Masanori Sasatsu; Takayoshi Suzuki; Masashi Matsushima; Jun Koike; Muneki Igarashi; Hideki Ozawa; Ryuki Fukuda; Atsushi Takagi

Disclosures

J Gastroenterol Hepatol. 2012;27(5):888-892. 

In This Article

Methods

Patients

A total of 229 patients diagnosed with an H. pylori infection participated in this study from April 2008 to August 2010. Patients were defined as positive for H. pylori infection if the culture was positive, or if histology and rapid urease test were positive. The following exclusion criteria were applied: age below 18 or above 80 years, previous H. pylori eradication, and the use of antimicrobials or gastrointestinal medications like PPI within the previous 2 months. The ethics committee of Tokai University Hospital approved the protocol, and written informed consent was obtained from all patients.

Determination of CAM Resistance

CAM resistance was determined by the detection of a mutation in 23S rRNA using nested polymerase chain reaction (PCR) and direct sequencing of DNA from pretreatment feces.[12] Briefly, pretreatment feces were pooled and kept in −20°C. DNA was extracted from feces using a bead-crushing method. Approximately 50 mg of feces was added to a tube with sodium phosphate buffer and 7.5 M guanidine solution and homogenized. The solution was centrifuged and subject to nested PCR. Nested PCR for the detection of mutations in the H. pylori 23S rRNA gene was performed. PCR primer pairs are shown in Table 1. DNA sequencing was carried out using an ABI PRISM 3100 DNA sequencer (Applied Biosystems, Carlsbad, CA, USA).

Yogurt

The yogurt containing L. gasseri OLL2716 (≥ 109c.f.u.) was obtained from Meiji Dairies Corporation, Tokyo, Japan

Protocol

A total of 229 patients were randomized either to 1 week of triple therapy comprising rabeprazole (10 mg bid), amoxicillin (750 mg bid), and clarithromycin (200 mg bid) or triple therapy plus L. gasseri-containing yogurt. In the yogurt-plus-triple therapy group, yogurt containing L. gasseri OLL2716 (112 g) was consumed twice daily for 4 weeks (3 weeks pretreatment followed by 1 week during eradication therapy). The triple therapy regimen in this study used the same doses based on a large-scale study in Japan.[13] Randomization was carried out according to a computer-generated randomization list. As a combination of the urea breath test (UBT) and a stool antigen test is useful for the clinical evaluation of eradication therapy,[14] H. pylori eradication was diagnosed based on UBT and stool antigen test after 8 weeks of eradication. For discordant results between UBT and stool antigen test, endoscopic biopsy specimens were obtained and H. pylori culture was carried out.

13C-urea Breath Test

The urea breath test was conducted 8 weeks after the end of eradication therapy. It was performed after overnight fasting using 100 mg 13C-urea tablets (Dainippon Sumitomo Pharmaceutical Co., Tokyo, Japan). Breath samples were collected before and 20 min after the ingestion of 13C-urea. The cut-off value of 13C value was defined as 3.5‰.[15]

H. Pylori Stool Antigen Test

Stool samples were frozen at −70°C until assaying. The investigators were blinded to the results of other H. pylori tests. A commercial EIA (Testmate H. pylori; Wakamoto Pharmaceutical, Tokyo, Japan) was used to detect H. pylori, according to the manufacturer's instructions. About a 100-mg fecal sample was diluted in 0.4 mL of dilution buffer. Fifty microliters of diluted fecal sample and peroxidase-conjugated monoclonal antibody were added to each well of microtiter plates, and the plates were incubated for 1 h at 25°C. The absorbance at dual wavelengths (450 and 630 nm) was measured on a microplate reader. The cut-off value for the stool antigen test was < 0.100 for a negative and ≥ 0.100 for a positive result.[15]

Statistical Analysis

The number of patients required for the study was calculated such that a difference of 17% in the eradication rates between the two study groups could be detected. Thus we calculated that at least 110 patients per group were required to give the study 80% power at a significant level of 5%. Eradication rates were determined for both groups employing per-protocol (PP) and intention-to-treat (ITT) analyses. For ITT analysis, all enrolled patients were included. For PP analysis, those who had not undergone the UBT and stool antigen test, or those who had taken less than 70% of any drugs, were excluded (Fig. 1).

Figure 1.

Flow schematic of the study involving intention-to-treat (ITT) and per-protocol (PP) analyses.

Eradication rates were compared using the χ2-test. Significance was set at P < 0.05. Statistical analysis was performed using spss 17.0J Windows (spss Japan, Inc., Tokyo, Japan).

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