Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis
Braz. j. infect. dis
;
15(3): 204-210, May-June 2011. ilus, tab
Article
in English
| LILACS
| ID: lil-589949
ABSTRACT
OBJECTIVE:
The objective of this study was to analyze different primers that are commonly used in epidemiological studies for the detection of Leishmania DNA by PCR, and to compare them to the conventional direct parasite search for American cutaneous leishmaniasis (ACL) diagnosis. MATERIAL ANDMETHODS:
Five pairs of primers, four of them derived from Leishmania kDNA sequences (MP3H-MP1L; B1-B2; LBF1-LBR1; 13A-13B), and one derived from the SL RNA (mini-exon) gene repeat (LU5A-LB3C), reported previously, were used.RESULTS:
The MP3H-MP1L primers were the best at amplifying the DNA, detecting 2 fg of Leishmania spp. DNA. The 13A-13B primers presented the worst performance, detecting 512 x 10³ fg of DNA.CONCLUSION:
The wide variation in the analytical sensitivity of the primers used in the PCR, and the significant differences from the conventional method of ACL diagnosis found in this study, emphasize the importance of standardizing the PCR technique, analyzing sensitivity, and selecting suitable oligonucleotide primers.
Full text:
Available
Index:
LILACS (Americas)
Main subject:
DNA, Protozoan
/
Leishmaniasis, Cutaneous
/
DNA Primers
/
Leishmania
Type of study:
Diagnostic study
Limits:
Animals
/
Humans
Language:
English
Journal:
Braz. j. infect. dis
Journal subject:
Communicable Diseases
Year:
2011
Type:
Article
/
Project document
Affiliation country:
Brazil
Institution/Affiliation country:
Universidade Estadual de Maringá/BR
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