Detection of Helicobacter pylori in paraffin-embedded gastric biopsy specimens by in situ hybridization

Tuomo J. Karttunen, Robert M. Genta, Boris Yoffe, Charles Y. Hachem, David Y. Graham, Fouad A K El-Zaatari

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Helicobacter pylori is the causative agent of chronic gastritis, peptic ulcers, and is also associated with gastric cancer. Eradication of H pylori infection has proven to be difficult to confirm. The authors developed a nonradioactive in situ hybridization method for detection of H pylori and compared it with conventional methods for diagnosis of the infection. Reverse transcription followed by polymerase chain reaction (PCR) with two 22-base primers was used to amplify a 520 base pair (bp) segment of 16S rRNA from H pylori. The PCR product was labeled with digoxigenin and used as a probe. Specificity of the probe was tested by dot blot hybridization against DNA from 30 different strains of related and unrelated bacteria. Specificity of in situ hybridization assay was proven by the lack of hybridization on sections with gram negative and positive bacteria other than H pylori, with an unrelated probe, without probe, and after RNase treatment. A random sample of 15 biopsy specimens was blindly studied by in situ hybridization method and the results were compared with those of culture and conventional histology. Comparison of in situ hybridization and conventional histology showed agreement in all 15 specimens (5 negative and 10 positive). Between culture and in situ hybridization there was agreement in 13 cases (8 positive, 5 negative). Two cases were negative by culture but positive by in situ hybridization and histology. Nonradioactive in situ hybridization provides a sensitive and specific method for detection of H pylori infection. It should be particularly useful for confirmation of infection in cases equivocal with other methods, and potentially useful in post-treatment evaluation.

Original languageEnglish (US)
Pages (from-to)305-311
Number of pages7
JournalAmerican Journal of Clinical Pathology
Volume106
Issue number3
StatePublished - Sep 1996

Fingerprint

Helicobacter pylori
Paraffin
In Situ Hybridization
Stomach
Pylorus
Biopsy
Histology
Infection
Digoxigenin
Polymerase Chain Reaction
Gram-Positive Bacteria
Gastritis
Ribonucleases
Gram-Negative Bacteria
Peptic Ulcer
Base Pairing
Reverse Transcription
Stomach Neoplasms
Bacteria
DNA

Keywords

  • Gastritis
  • Helicobacter pylori
  • In situ hybridization

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Karttunen, T. J., Genta, R. M., Yoffe, B., Hachem, C. Y., Graham, D. Y., & El-Zaatari, F. A. K. (1996). Detection of Helicobacter pylori in paraffin-embedded gastric biopsy specimens by in situ hybridization. American Journal of Clinical Pathology, 106(3), 305-311.

Detection of Helicobacter pylori in paraffin-embedded gastric biopsy specimens by in situ hybridization. / Karttunen, Tuomo J.; Genta, Robert M.; Yoffe, Boris; Hachem, Charles Y.; Graham, David Y.; El-Zaatari, Fouad A K.

In: American Journal of Clinical Pathology, Vol. 106, No. 3, 09.1996, p. 305-311.

Research output: Contribution to journalArticle

Karttunen, TJ, Genta, RM, Yoffe, B, Hachem, CY, Graham, DY & El-Zaatari, FAK 1996, 'Detection of Helicobacter pylori in paraffin-embedded gastric biopsy specimens by in situ hybridization', American Journal of Clinical Pathology, vol. 106, no. 3, pp. 305-311.
Karttunen TJ, Genta RM, Yoffe B, Hachem CY, Graham DY, El-Zaatari FAK. Detection of Helicobacter pylori in paraffin-embedded gastric biopsy specimens by in situ hybridization. American Journal of Clinical Pathology. 1996 Sep;106(3):305-311.
Karttunen, Tuomo J. ; Genta, Robert M. ; Yoffe, Boris ; Hachem, Charles Y. ; Graham, David Y. ; El-Zaatari, Fouad A K. / Detection of Helicobacter pylori in paraffin-embedded gastric biopsy specimens by in situ hybridization. In: American Journal of Clinical Pathology. 1996 ; Vol. 106, No. 3. pp. 305-311.
@article{1955c36e92f44035bed22f79a32a566e,
title = "Detection of Helicobacter pylori in paraffin-embedded gastric biopsy specimens by in situ hybridization",
abstract = "Helicobacter pylori is the causative agent of chronic gastritis, peptic ulcers, and is also associated with gastric cancer. Eradication of H pylori infection has proven to be difficult to confirm. The authors developed a nonradioactive in situ hybridization method for detection of H pylori and compared it with conventional methods for diagnosis of the infection. Reverse transcription followed by polymerase chain reaction (PCR) with two 22-base primers was used to amplify a 520 base pair (bp) segment of 16S rRNA from H pylori. The PCR product was labeled with digoxigenin and used as a probe. Specificity of the probe was tested by dot blot hybridization against DNA from 30 different strains of related and unrelated bacteria. Specificity of in situ hybridization assay was proven by the lack of hybridization on sections with gram negative and positive bacteria other than H pylori, with an unrelated probe, without probe, and after RNase treatment. A random sample of 15 biopsy specimens was blindly studied by in situ hybridization method and the results were compared with those of culture and conventional histology. Comparison of in situ hybridization and conventional histology showed agreement in all 15 specimens (5 negative and 10 positive). Between culture and in situ hybridization there was agreement in 13 cases (8 positive, 5 negative). Two cases were negative by culture but positive by in situ hybridization and histology. Nonradioactive in situ hybridization provides a sensitive and specific method for detection of H pylori infection. It should be particularly useful for confirmation of infection in cases equivocal with other methods, and potentially useful in post-treatment evaluation.",
keywords = "Gastritis, Helicobacter pylori, In situ hybridization",
author = "Karttunen, {Tuomo J.} and Genta, {Robert M.} and Boris Yoffe and Hachem, {Charles Y.} and Graham, {David Y.} and El-Zaatari, {Fouad A K}",
year = "1996",
month = "9",
language = "English (US)",
volume = "106",
pages = "305--311",
journal = "American Journal of Clinical Pathology",
issn = "0002-9173",
publisher = "American Society of Clinical Pathologists",
number = "3",

}

TY - JOUR

T1 - Detection of Helicobacter pylori in paraffin-embedded gastric biopsy specimens by in situ hybridization

AU - Karttunen, Tuomo J.

AU - Genta, Robert M.

AU - Yoffe, Boris

AU - Hachem, Charles Y.

AU - Graham, David Y.

AU - El-Zaatari, Fouad A K

PY - 1996/9

Y1 - 1996/9

N2 - Helicobacter pylori is the causative agent of chronic gastritis, peptic ulcers, and is also associated with gastric cancer. Eradication of H pylori infection has proven to be difficult to confirm. The authors developed a nonradioactive in situ hybridization method for detection of H pylori and compared it with conventional methods for diagnosis of the infection. Reverse transcription followed by polymerase chain reaction (PCR) with two 22-base primers was used to amplify a 520 base pair (bp) segment of 16S rRNA from H pylori. The PCR product was labeled with digoxigenin and used as a probe. Specificity of the probe was tested by dot blot hybridization against DNA from 30 different strains of related and unrelated bacteria. Specificity of in situ hybridization assay was proven by the lack of hybridization on sections with gram negative and positive bacteria other than H pylori, with an unrelated probe, without probe, and after RNase treatment. A random sample of 15 biopsy specimens was blindly studied by in situ hybridization method and the results were compared with those of culture and conventional histology. Comparison of in situ hybridization and conventional histology showed agreement in all 15 specimens (5 negative and 10 positive). Between culture and in situ hybridization there was agreement in 13 cases (8 positive, 5 negative). Two cases were negative by culture but positive by in situ hybridization and histology. Nonradioactive in situ hybridization provides a sensitive and specific method for detection of H pylori infection. It should be particularly useful for confirmation of infection in cases equivocal with other methods, and potentially useful in post-treatment evaluation.

AB - Helicobacter pylori is the causative agent of chronic gastritis, peptic ulcers, and is also associated with gastric cancer. Eradication of H pylori infection has proven to be difficult to confirm. The authors developed a nonradioactive in situ hybridization method for detection of H pylori and compared it with conventional methods for diagnosis of the infection. Reverse transcription followed by polymerase chain reaction (PCR) with two 22-base primers was used to amplify a 520 base pair (bp) segment of 16S rRNA from H pylori. The PCR product was labeled with digoxigenin and used as a probe. Specificity of the probe was tested by dot blot hybridization against DNA from 30 different strains of related and unrelated bacteria. Specificity of in situ hybridization assay was proven by the lack of hybridization on sections with gram negative and positive bacteria other than H pylori, with an unrelated probe, without probe, and after RNase treatment. A random sample of 15 biopsy specimens was blindly studied by in situ hybridization method and the results were compared with those of culture and conventional histology. Comparison of in situ hybridization and conventional histology showed agreement in all 15 specimens (5 negative and 10 positive). Between culture and in situ hybridization there was agreement in 13 cases (8 positive, 5 negative). Two cases were negative by culture but positive by in situ hybridization and histology. Nonradioactive in situ hybridization provides a sensitive and specific method for detection of H pylori infection. It should be particularly useful for confirmation of infection in cases equivocal with other methods, and potentially useful in post-treatment evaluation.

KW - Gastritis

KW - Helicobacter pylori

KW - In situ hybridization

UR - http://www.scopus.com/inward/record.url?scp=0029758472&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029758472&partnerID=8YFLogxK

M3 - Article

C2 - 8816586

AN - SCOPUS:0029758472

VL - 106

SP - 305

EP - 311

JO - American Journal of Clinical Pathology

JF - American Journal of Clinical Pathology

SN - 0002-9173

IS - 3

ER -