Development and application of the polymerase chain reaction for the detection and identification of Leishmania parasites in clinical material.
Detection, diagnosis and identification of Leishmaniasis may be difficult owing to low numbers of parasites present in clinical samples. The PCR has improved the sensitivity and specificity of diagnosis of several infectious diseases. A leishmania specific PCR assay was developed based on the SSUrRNA genes which amplifies DNA of all Leishmania species. Point mutations occurring within the rRNA genes allow differentiation of the Leishmania complexes using primers constructed with the 3/ ends complementary to the specific point mutations present in the SSU rRNA genes of the Leishmania species. Biopsy material, blood, lesion impressions and blood spots on filter paper can be used in the assay. In a longitudinal study on the incidence rates of VL, subclinical cases and PKDL in an endemic region of Sudan, filter paper blood spots from proven and suspected VL patients, PKDL and control samples from an endemic region in Sudan are being taken. The blood spots were analyzed in the DAT and by PCR and results compared with clinical and parasitological data. The first results indicate that the PCR on blood spots is a simple and sensitive means of detecting active VL; in PKDL patients parasites are detectable in the skin.
Meredith
S E
SE
Royal Tropical Institute, Amsterdam, The Netherlands.
Zijlstra
E E
EE
Schoone
G J
GJ
Kroon
C C
CC
van Eys
G J
GJ
Schaeffer
K U
KU
el-Hassan
A M
AM
Lawyer
P G
PG
D000818:Animals; D001483:Base Sequence; D018105:DNA, Kinetoplast / Q000235:genetics; D005069:Evaluation Studies as Topic; D006801:Humans; D015994:Incidence; D007891:Leishmania / Q000145:classification / Q000235:genetics*; D007896:Leishmaniasis / Q000175:diagnosis / Q000453:epidemiology / Q000469:parasitology*; D008137:Longitudinal Studies; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D016053:RNA, Protozoan / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D012680:Sensitivity and Specificity; D013397:Sudan / Q000453:epidemiology
D018105:DNA, Kinetoplast; D016053:RNA, Protozoan; D012335:RNA, Ribosomal