Detection of borrelia burgdorferi in human blood and urine using the polymerase chain reaction

Byron S. McGuire, Francis W. Chandler, Michael W. Felz, Lee O. Huey, Richard S. Field

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

We investigated the use of the polymerase chain reaction (PCR) to detect Borrelia burgdorferi strain B-31 in human blood and urine experimentally inoculated with 5 and 1 borreliae/cm3, respectively, and to biotinylate a DNA probe specific for B. burgdorferi in the dot blot and Southern blot assays. When the blood and urine samples were subjected to PCR, a 370-bp amplified product was consistently visible on agarose gel electrophoresis after 30 and 45 cycles, respectively. The total human genomic DNA extracted from a l-cm3 sample of inoculated blood was approximately 6.25 μg, and the total amount of B. burgdorferi DNA was estimated to be 0.01 pg/6.25 μg of the human DNA. For PCR. 2.5 μg of human DNA which contained the equivalent of 0.004 pg of borrelia DNA (approximately two borreliae) were used for enzymatic amplification. When 1/20 or 1/10 of the PCR-amplificd products were used either for dot blot or Southern blot hybridization, the accessible copies of amplified B. burgdorferi DNA were sufficient for detectable hybridization to occur. PCR amplification of B. burgdorferi DNA in clinical specimens followed by dot blot hybridization may be a valuable adjunct or alternative to current but inadequate laboratory methods for the diagnosis of Lyme disease.

Original languageEnglish (US)
Pages (from-to)163-167
Number of pages5
JournalPathobiology
Volume60
Issue number3
DOIs
StatePublished - Jan 1 1992

Fingerprint

Borrelia burgdorferi
Urine
Polymerase Chain Reaction
Borrelia
DNA
Southern Blotting
Lyme Disease
Agar Gel Electrophoresis
Clinical Laboratory Techniques
DNA Probes

Keywords

  • Blood
  • Borrelia burgdorferi
  • Experimental infection
  • Lyme disease
  • Polymerase chain reaction
  • Urine

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

Cite this

McGuire, B. S., Chandler, F. W., Felz, M. W., Huey, L. O., & Field, R. S. (1992). Detection of borrelia burgdorferi in human blood and urine using the polymerase chain reaction. Pathobiology, 60(3), 163-167. https://doi.org/10.1159/000163717

Detection of borrelia burgdorferi in human blood and urine using the polymerase chain reaction. / McGuire, Byron S.; Chandler, Francis W.; Felz, Michael W.; Huey, Lee O.; Field, Richard S.

In: Pathobiology, Vol. 60, No. 3, 01.01.1992, p. 163-167.

Research output: Contribution to journalArticle

McGuire, BS, Chandler, FW, Felz, MW, Huey, LO & Field, RS 1992, 'Detection of borrelia burgdorferi in human blood and urine using the polymerase chain reaction', Pathobiology, vol. 60, no. 3, pp. 163-167. https://doi.org/10.1159/000163717
McGuire, Byron S. ; Chandler, Francis W. ; Felz, Michael W. ; Huey, Lee O. ; Field, Richard S. / Detection of borrelia burgdorferi in human blood and urine using the polymerase chain reaction. In: Pathobiology. 1992 ; Vol. 60, No. 3. pp. 163-167.
@article{358bb4512ed24951a02f8e58817aa8a7,
title = "Detection of borrelia burgdorferi in human blood and urine using the polymerase chain reaction",
abstract = "We investigated the use of the polymerase chain reaction (PCR) to detect Borrelia burgdorferi strain B-31 in human blood and urine experimentally inoculated with 5 and 1 borreliae/cm3, respectively, and to biotinylate a DNA probe specific for B. burgdorferi in the dot blot and Southern blot assays. When the blood and urine samples were subjected to PCR, a 370-bp amplified product was consistently visible on agarose gel electrophoresis after 30 and 45 cycles, respectively. The total human genomic DNA extracted from a l-cm3 sample of inoculated blood was approximately 6.25 μg, and the total amount of B. burgdorferi DNA was estimated to be 0.01 pg/6.25 μg of the human DNA. For PCR. 2.5 μg of human DNA which contained the equivalent of 0.004 pg of borrelia DNA (approximately two borreliae) were used for enzymatic amplification. When 1/20 or 1/10 of the PCR-amplificd products were used either for dot blot or Southern blot hybridization, the accessible copies of amplified B. burgdorferi DNA were sufficient for detectable hybridization to occur. PCR amplification of B. burgdorferi DNA in clinical specimens followed by dot blot hybridization may be a valuable adjunct or alternative to current but inadequate laboratory methods for the diagnosis of Lyme disease.",
keywords = "Blood, Borrelia burgdorferi, Experimental infection, Lyme disease, Polymerase chain reaction, Urine",
author = "McGuire, {Byron S.} and Chandler, {Francis W.} and Felz, {Michael W.} and Huey, {Lee O.} and Field, {Richard S.}",
year = "1992",
month = "1",
day = "1",
doi = "10.1159/000163717",
language = "English (US)",
volume = "60",
pages = "163--167",
journal = "Pathobiology",
issn = "1015-2008",
publisher = "S. Karger AG",
number = "3",

}

TY - JOUR

T1 - Detection of borrelia burgdorferi in human blood and urine using the polymerase chain reaction

AU - McGuire, Byron S.

AU - Chandler, Francis W.

AU - Felz, Michael W.

AU - Huey, Lee O.

AU - Field, Richard S.

PY - 1992/1/1

Y1 - 1992/1/1

N2 - We investigated the use of the polymerase chain reaction (PCR) to detect Borrelia burgdorferi strain B-31 in human blood and urine experimentally inoculated with 5 and 1 borreliae/cm3, respectively, and to biotinylate a DNA probe specific for B. burgdorferi in the dot blot and Southern blot assays. When the blood and urine samples were subjected to PCR, a 370-bp amplified product was consistently visible on agarose gel electrophoresis after 30 and 45 cycles, respectively. The total human genomic DNA extracted from a l-cm3 sample of inoculated blood was approximately 6.25 μg, and the total amount of B. burgdorferi DNA was estimated to be 0.01 pg/6.25 μg of the human DNA. For PCR. 2.5 μg of human DNA which contained the equivalent of 0.004 pg of borrelia DNA (approximately two borreliae) were used for enzymatic amplification. When 1/20 or 1/10 of the PCR-amplificd products were used either for dot blot or Southern blot hybridization, the accessible copies of amplified B. burgdorferi DNA were sufficient for detectable hybridization to occur. PCR amplification of B. burgdorferi DNA in clinical specimens followed by dot blot hybridization may be a valuable adjunct or alternative to current but inadequate laboratory methods for the diagnosis of Lyme disease.

AB - We investigated the use of the polymerase chain reaction (PCR) to detect Borrelia burgdorferi strain B-31 in human blood and urine experimentally inoculated with 5 and 1 borreliae/cm3, respectively, and to biotinylate a DNA probe specific for B. burgdorferi in the dot blot and Southern blot assays. When the blood and urine samples were subjected to PCR, a 370-bp amplified product was consistently visible on agarose gel electrophoresis after 30 and 45 cycles, respectively. The total human genomic DNA extracted from a l-cm3 sample of inoculated blood was approximately 6.25 μg, and the total amount of B. burgdorferi DNA was estimated to be 0.01 pg/6.25 μg of the human DNA. For PCR. 2.5 μg of human DNA which contained the equivalent of 0.004 pg of borrelia DNA (approximately two borreliae) were used for enzymatic amplification. When 1/20 or 1/10 of the PCR-amplificd products were used either for dot blot or Southern blot hybridization, the accessible copies of amplified B. burgdorferi DNA were sufficient for detectable hybridization to occur. PCR amplification of B. burgdorferi DNA in clinical specimens followed by dot blot hybridization may be a valuable adjunct or alternative to current but inadequate laboratory methods for the diagnosis of Lyme disease.

KW - Blood

KW - Borrelia burgdorferi

KW - Experimental infection

KW - Lyme disease

KW - Polymerase chain reaction

KW - Urine

UR - http://www.scopus.com/inward/record.url?scp=0026587120&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026587120&partnerID=8YFLogxK

U2 - 10.1159/000163717

DO - 10.1159/000163717

M3 - Article

C2 - 1627262

AN - SCOPUS:0026587120

VL - 60

SP - 163

EP - 167

JO - Pathobiology

JF - Pathobiology

SN - 1015-2008

IS - 3

ER -