The Helicobacter pylori L-form: formation and isolation in the human bile cultures in vitro and in the gallbladders of patients with biliary diseases.
BACKGROUND
The Helicobacter pylori is considered the important causative agent causing biliary diseases, but the H. pylori can be isolated from very few gallbladder specimens with diseases. We studied the formation of H. pylori L-forms in bile in vitro and isolated the H. pylori L-forms from gallbladder of patients with biliary diseases.


METHODS
We inoculated the H. pylori into the human bile to induce the L-form in vitro. The gallbladder specimens were collected from patients with biliary diseases to isolate the bacterial L-forms by the nonhigh osmotic isolation technique, and the H. pylori L-forms in the L-form isolates were identified by the gene assay for the H. pylori-specific genes 16S rRNA and UreA.


RESULTS
The H. Pylori cannot be isolated from the bile-induced cultures, but the H. pylori L-form can be isolated from the H. pylori-negative bile-induced cultures. The L-form isolates of bile-induced cultures showed a positive reaction of the H. pylori-specific genes by PCR, and the coincidence ratio of the nucleotide sequences between the L-forms and the H. pylori is 99%. The isolation rate of bacteria L-form is 93.2% in the gallbladder specimens with bacteria-negative isolation culture by the nonhigh osmotic isolation technique, and the positive rate of the H. pylori-specific genes in the L-form isolates is 7.1% in the bacterial L-form-positive isolation cultures by the PCR.


CONCLUSIONS
H. pylori can be rapidly induced into the L-form in the human bile; the L-form, as the latent bacteria, can live in the host gallbladder for a long times, and they made the host became a latent carrier of the H. pylori L-form. The H. pylori L-form can be isolated by the nonhigh osmotic isolation technique, and the variant can be identified by the gene assay for the H. pylori-specific genes 16S rRNA and reA.
Department of Medical Microbiology and Parasitology, Institutes of Medical Science, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Dan N
DN
Wang
Wen J
WJ
Ding
Yao Z
YZ
Pan
Ke L
KL
Tang
Tao
T
Wang
Xiao L
XL
She
He
H
Wang
D001646:Bile / Q000382:microbiology*; D001660:Biliary Tract Diseases / Q000382:microbiology*; D002353:Carrier State / Q000382:microbiology; D005704:Gallbladder / Q000382:microbiology*; D016481:Helicobacter Infections / Q000382:microbiology*; D016480:Helicobacter pylori / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007740:L Forms / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D000091402:Prohibitins; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D014510:Urease / Q000235:genetics
C000717331:PHB2 protein, human; D000091402:Prohibitins; D012336:RNA, Ribosomal, 16S; D014510:Urease
Bacterial L-form; Helicobacter pylori; biliary diseases; epidemiology; evaluation