Growth Medium Containing Cyclodextrin and Low Concentration of Horse Serum for Cultivation of <i>Helicobacter pylori</i>

Morshed, Muhammad; Karita, Mikio; Konishi, Hisanori; Okita, Kiwamu; Nakazawa, Takahiro

doi:10.1111/j.1348-0421.1994.tb02143.x

Cited by 15 publications

(12 citation statements)

References 13 publications

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“…As in our study, several other groups have reported growth enhancement of H. pylori when culture medium is supplemented with ␤-cyclodextrin (10,16,22,24). However, it appears that not all preparations of ␤-cyclodextrins are able to support H. pylori growth (8,16), perhaps explaining why one group was unable to obtain growth of H. pylori in media supplemented with ␤-cyclodextrin (30).…”

Section: Discussionsupporting

confidence: 65%

Helicobacter pylori Growth and Urease Detection in the Chemically Defined Medium Ham's F-12 Nutrient Mixture

2001

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Obstacles continue to hinder in vitro studies of the gastric human pathogen Helicobacter pylori, including difficulty culturing the organism in the absence of serum or blood, rapid loss of viability following exponential growth due to autolysis, and the necessity for using high starting inocula. We demonstrate that H. pylori grows in the chemically defined broth medium Ham's F-12 nutrient mixture (F-12) in the absence of fetal bovine serum (FBS); this represents a breakthrough for studies in which serum components or proteins interfere with interpretation of results. Cultures can be continually passaged in fresh, FBS-free F-12 medium at an initial inoculum of only ϳ10 3 CFU/ml. All H. pylori strains (n ‫؍‬ 21), including fresh clinical isolates, grew in serum-free F-12. H. pylori grew poorly in the related medium, F-10, unless additional zinc was supplied. Enhanced growth of H. pylori in F-12 broth was obtained by addition of bovine serum albumin (BSA) (1 mg/ml), ␤-cyclodextrin (200 g/ml), or cholesterol (50 g/ml). H. pylori also grew in several simplified versions of F-12 broth lacking glucose and most vitamins but containing hypoxanthine, pyruvate, and all 20 amino acids. On F-12 medium solidified with agar, H. pylori only grew when BSA (98% pure; 1 mg/ml), cholesterol (50 g/ml), ␤-cyclodextrin (200 g/ml), or FBS (2 to 4%) was added; addition of urea and phenol allowed colorimetric detection of urease activity. Thus, F-12 agar plus cholesterol or ␤-cyclodextrin represents the first transparent chemically defined agar and the first urease indicator agar for H. pylori. Several lines of evidence suggested that BSA itself is not responsible for H. pylori growth enhancement in F-12 containing BSA or FBS. Taken together, these innovations represent significant advances in the cultivation and recovery of H. pylori using chemically defined media. Use of F-12 or its derivatives may lead to improved understanding of H. pylori metabolism, virulence factors, and transmission, and result in improved recovery and identification of H. pylori from clinical specimens.

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Section: Discussionsupporting

confidence: 65%

Helicobacter pylori Growth and Urease Detection in the Chemically Defined Medium Ham's F-12 Nutrient Mixture

2001

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“…Other bacterially mediated autoimmune diseases in which persistent infection and induction of autoimmune pathology have been postulated to contribute to disease pathology include gastric inflammation (Helicobacter pylori) (4,44) and arteriosclerosis (Chlamydia pneumoniae) (22,36). In the former case, the organism is clearly implicated, since the disease responds to elimination of the organism, whereas in the latter case, the organism may either contribute to the pathogenesis of the atherosclerotic plaque or merely act as a colonizer of the diseased and compromised tissue.…”

Section: Discussionmentioning

confidence: 99%

Reverse Transcriptase-PCR Analysis of Bacterial rRNA for Detection and Characterization of Bacterial Species in Arthritis Synovial Tissue

Kempsell¹,

Cox

Hurle³

et al. 2000

Infect Immun

119

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Onset of rheumatoid arthritis (RA) is widely believed to be preceded by exposure to some environmental trigger such as bacterial infectious agents. The influence of bacteria on RA disease onset or pathology has to date been controversial, due to inconsistencies between groups in the report of bacterial species isolated from RA disease tissue. Using a modified technique of reverse transcriptase-PCR amplification, we have detected bacterial rRNA in the synovial tissue of late-stage RA and non-RA arthritis controls. This may be suggestive of the presence of live bacteria. Sequencing of cloned complementary rDNA (crDNA) products revealed a number of bacterial sequences in joint tissue from each patient, and from these analyses a comprehensive profile of the organisms present was compiled. This revealed a number of different organisms in each patient, some of which are common to both RA and non-RA controls and are probably opportunistic colonizers of previously diseased tissue and others which are unique species. These latter organisms may be candidates for a specific role in disease pathology and require further investigation to exclude them as causative agents in the complex bacterial millieu. In addition, many of the detected bacterial species have not been identified previously from synovial tissue or fluid from arthritis patients. These may not be easily cultivable, since they were not revealed in previous studies using conventional in vitro bacterial culture methods. In situ hybridization analyses have revealed the joint-associated bacterial rRNA to be both intra- and extracellular. The role of viable bacteria or their nucleic acids as triggers in disease onset or pathology in either RA or non-RA arthritis controls is unclear and requires further investigation.

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“…In an effort to overcome some of these problems, various studies have described the use of chemically defined compounds belonging to the CD group as substitutes for serum in both solid and liquid media [6,7,15,21]. CDs are cyclic oligosaccharides that are thought to reduce the effective free concentration of toxic substrates and ⁄ or products to levels that are not inhibitory to microorganisms (reviewed in [32]).…”

Section: Discussionmentioning

confidence: 99%

The Use of AlbuMAX II^® as a Blood or Serum Alternative for the Culture of Helicobacter pylori

2012

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Growth Medium Containing Cyclodextrin and Low Concentration of Horse Serum for Cultivation of Helicobacter pylori

Abstract: Abstract:The

Cited by 15 publications

References 13 publications

Helicobacter pylori Growth and Urease Detection in the Chemically Defined Medium Ham's F-12 Nutrient Mixture

Helicobacter pylori Growth and Urease Detection in the Chemically Defined Medium Ham's F-12 Nutrient Mixture

Reverse Transcriptase-PCR Analysis of Bacterial rRNA for Detection and Characterization of Bacterial Species in Arthritis Synovial Tissue

The Use of AlbuMAX II^® as a Blood or Serum Alternative for the Culture of Helicobacter pylori

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Growth Medium Containing Cyclodextrin and Low Concentration of Horse Serum for Cultivation of Helicobacter pylori

Abstract: Abstract:The

Cited by 15 publications

References 13 publications

Helicobacter pylori Growth and Urease Detection in the Chemically Defined Medium Ham's F-12 Nutrient Mixture

Helicobacter pylori Growth and Urease Detection in the Chemically Defined Medium Ham's F-12 Nutrient Mixture

Reverse Transcriptase-PCR Analysis of Bacterial rRNA for Detection and Characterization of Bacterial Species in Arthritis Synovial Tissue

The Use of AlbuMAX II® as a Blood or Serum Alternative for the Culture of Helicobacter pylori

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Product

Resources

About

The Use of AlbuMAX II^® as a Blood or Serum Alternative for the Culture of Helicobacter pylori