Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues

Aleco D'Andrea, Yarelys Zamora Martinez, Rosa Alduina, Vincenzo Monteverde, Carmen Fernandez Molina, Maria Vitale

Research output: Contribution to journalArticle

Abstract

In this study we compared two polymerase chain reaction (PCR) methods using either 16S ribosomal RNA (rRNA) or 23S rRNA gene primers for the detection of different Leptospira interrogans serovars. The performance of these two methods was assessed using DNA extracted from bovine tissues previously inoculated with several bacterial suspensions. PCR was performed on the same tissues before and after the formalin-fixed, paraffin-embedding procedure (FFPE tissues). The 23S rDNA PCR detected all fresh and FFPE positive tissues while the 16S rDNA-based protocol detected primarily the positive fresh tissues. Both methods are specific for pathogenic L. interrogans. The 23S-based PCR method successfully detected Leptospira in four dubious cases of human leptospirosis from archival tissue specimens and one leptospirosis-positive canine specimen. A sensitive method for leptospirosis identification in FFPE tissues would be a useful tool to screen histological specimen archives and gain a better assessment of human leptospirosis prevalence, especially in tropical countries, where large outbreaks can occur following the rainy season.

Original languageEnglish
Pages (from-to)85-88
Number of pages4
JournalMemorias do Instituto Oswaldo Cruz
Volume107
Issue number1
Publication statusPublished - Feb 2012

Fingerprint

Leptospira interrogans
Paraffin
Formaldehyde
Leptospirosis
Polymerase Chain Reaction
Ribosomal DNA
23S Ribosomal RNA
Paraffin Embedding
Leptospira
rRNA Genes
Disease Outbreaks
Canidae
Suspensions
DNA

Keywords

  • Diagnostics
  • FFPE tissue
  • Leptospira
  • PCR

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

D'Andrea, A., Martinez, Y. Z., Alduina, R., Monteverde, V., Molina, C. F., & Vitale, M. (2012). Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues. Memorias do Instituto Oswaldo Cruz, 107(1), 85-88.

Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues. / D'Andrea, Aleco; Martinez, Yarelys Zamora; Alduina, Rosa; Monteverde, Vincenzo; Molina, Carmen Fernandez; Vitale, Maria.

In: Memorias do Instituto Oswaldo Cruz, Vol. 107, No. 1, 02.2012, p. 85-88.

Research output: Contribution to journalArticle

D'Andrea, A, Martinez, YZ, Alduina, R, Monteverde, V, Molina, CF & Vitale, M 2012, 'Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues', Memorias do Instituto Oswaldo Cruz, vol. 107, no. 1, pp. 85-88.
D'Andrea A, Martinez YZ, Alduina R, Monteverde V, Molina CF, Vitale M. Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues. Memorias do Instituto Oswaldo Cruz. 2012 Feb;107(1):85-88.
D'Andrea, Aleco ; Martinez, Yarelys Zamora ; Alduina, Rosa ; Monteverde, Vincenzo ; Molina, Carmen Fernandez ; Vitale, Maria. / Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues. In: Memorias do Instituto Oswaldo Cruz. 2012 ; Vol. 107, No. 1. pp. 85-88.
@article{d9e7c8168aca48798b17620e7f8de51e,
title = "Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues",
abstract = "In this study we compared two polymerase chain reaction (PCR) methods using either 16S ribosomal RNA (rRNA) or 23S rRNA gene primers for the detection of different Leptospira interrogans serovars. The performance of these two methods was assessed using DNA extracted from bovine tissues previously inoculated with several bacterial suspensions. PCR was performed on the same tissues before and after the formalin-fixed, paraffin-embedding procedure (FFPE tissues). The 23S rDNA PCR detected all fresh and FFPE positive tissues while the 16S rDNA-based protocol detected primarily the positive fresh tissues. Both methods are specific for pathogenic L. interrogans. The 23S-based PCR method successfully detected Leptospira in four dubious cases of human leptospirosis from archival tissue specimens and one leptospirosis-positive canine specimen. A sensitive method for leptospirosis identification in FFPE tissues would be a useful tool to screen histological specimen archives and gain a better assessment of human leptospirosis prevalence, especially in tropical countries, where large outbreaks can occur following the rainy season.",
keywords = "Diagnostics, FFPE tissue, Leptospira, PCR",
author = "Aleco D'Andrea and Martinez, {Yarelys Zamora} and Rosa Alduina and Vincenzo Monteverde and Molina, {Carmen Fernandez} and Maria Vitale",
year = "2012",
month = "2",
language = "English",
volume = "107",
pages = "85--88",
journal = "Memorias do Instituto Oswaldo Cruz",
issn = "0074-0276",
publisher = "Fundacao Oswaldo Cruz",
number = "1",

}

TY - JOUR

T1 - Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues

AU - D'Andrea, Aleco

AU - Martinez, Yarelys Zamora

AU - Alduina, Rosa

AU - Monteverde, Vincenzo

AU - Molina, Carmen Fernandez

AU - Vitale, Maria

PY - 2012/2

Y1 - 2012/2

N2 - In this study we compared two polymerase chain reaction (PCR) methods using either 16S ribosomal RNA (rRNA) or 23S rRNA gene primers for the detection of different Leptospira interrogans serovars. The performance of these two methods was assessed using DNA extracted from bovine tissues previously inoculated with several bacterial suspensions. PCR was performed on the same tissues before and after the formalin-fixed, paraffin-embedding procedure (FFPE tissues). The 23S rDNA PCR detected all fresh and FFPE positive tissues while the 16S rDNA-based protocol detected primarily the positive fresh tissues. Both methods are specific for pathogenic L. interrogans. The 23S-based PCR method successfully detected Leptospira in four dubious cases of human leptospirosis from archival tissue specimens and one leptospirosis-positive canine specimen. A sensitive method for leptospirosis identification in FFPE tissues would be a useful tool to screen histological specimen archives and gain a better assessment of human leptospirosis prevalence, especially in tropical countries, where large outbreaks can occur following the rainy season.

AB - In this study we compared two polymerase chain reaction (PCR) methods using either 16S ribosomal RNA (rRNA) or 23S rRNA gene primers for the detection of different Leptospira interrogans serovars. The performance of these two methods was assessed using DNA extracted from bovine tissues previously inoculated with several bacterial suspensions. PCR was performed on the same tissues before and after the formalin-fixed, paraffin-embedding procedure (FFPE tissues). The 23S rDNA PCR detected all fresh and FFPE positive tissues while the 16S rDNA-based protocol detected primarily the positive fresh tissues. Both methods are specific for pathogenic L. interrogans. The 23S-based PCR method successfully detected Leptospira in four dubious cases of human leptospirosis from archival tissue specimens and one leptospirosis-positive canine specimen. A sensitive method for leptospirosis identification in FFPE tissues would be a useful tool to screen histological specimen archives and gain a better assessment of human leptospirosis prevalence, especially in tropical countries, where large outbreaks can occur following the rainy season.

KW - Diagnostics

KW - FFPE tissue

KW - Leptospira

KW - PCR

UR - http://www.scopus.com/inward/record.url?scp=84857081573&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84857081573&partnerID=8YFLogxK

M3 - Article

C2 - 22310540

AN - SCOPUS:84857081573

VL - 107

SP - 85

EP - 88

JO - Memorias do Instituto Oswaldo Cruz

JF - Memorias do Instituto Oswaldo Cruz

SN - 0074-0276

IS - 1

ER -