Development of a rapid PCR method for identification of Helicobacter pylori in dental plaque and gastric biopsy specimens.
A rapid and simple polymerase chain reaction (PCR) method was developed to detect Helicobacter pylori in gastric biopsy specimens and dental plaque samples. The primers were targeted to the 16S rRNA sequence of Helicobacter pylori strain ATCC 43504. The system was found to have a theoretical detection level of 0.5 to 5 Helicobacter pylori cells in a 5 microliters sample of dental plaque. In the absence of plaque, the detection level was even better: theoretically, 0.05 to 0.5 Helicobacter pylori cells were detected in water suspension. However, this appeared to be due to the presence of free bacterial DNA in the culture used for the sensitivity determination. Thus, the actual sensitivity of the system was found to be fewer than five Helicobacter pylori cells, irrespective of the type of sample used. The method was then used to analyse 29 dental plaque and gastric biopsy specimens collected from patients with a history of recurrent peptic ulcer disease. Fourteen stomach specimens were positive for Helicobacter pylori when tested with the PCR method, while the respective figures with culture, histological examination and the urease test were 11, 12 and 9. No positive dental plaque samples were observed.
Department of Biochemistry and Biotechnology, A.I. Virtanen Institute, Kuopio, Finland.
J
J
Wahlfors
J H
JH
Meurman
J
J
Toskala
A
A
Korhonen
P
P
Alakuijala
E
E
Janatuinen
U M
UM
Kärkkkäinen
P
P
Nuutinen
J
J
Jänne
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D001706:Biopsy; D003773:Dental Plaque / Q000382:microbiology*; D005260:Female; D016480:Helicobacter pylori / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D013270:Stomach / Q000382:microbiology*