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Biomedical subjects

D C Barker

Publications and source records attributed to D C Barker.

81 records · Page 5Linked to original sources
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Leishmania (Viannia) lainsoni occupies a unique niche within the subgenus Viannia.

Leishmania lainsoni has recently been recognized as a new peripylarian species belonging to the subgenus Viannia and the L. braziliensis complex. It has been isolated from its sandfly vector, reservoir host and cutaneous lesions of human patients. Microscopical examination has shown characteristics which are different from those of other members of the L. braziliensis complex. Nuclear deoxyribonucleic acid (DNA) hybridization patterns with a beta tubulin probe and kinetoplast DNA buoyant density measurements show close similarities with other species of the L. braziliensis complex. However, kinetoplast DNA restriction enzyme fragment patterns of L. (V.) lainsoni isolates show similarities to L. mexicana complex species as well as weak cross hybridization. L. (V.) lainsoni is also amplified with L. braziliensis complex specific polymerase chain reaction (PCR) primers but it requires a lower annealing temperature and gives a 300 base pair PCR product. A possible model for the binding of PCR primers to the L. (V.) lainsoni kinetoplast DNA minicircle is proposed.

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Molecular tracking of infections by Leishmania infantum.

Leishmania infantum is a major opportunistic parasite in patients with acquired immune deficiency syndrome and is very variable in these subjects. Isoenzyme characterization is not able to explain this variability, since half of the stocks isolated from patients co-infected with human immunodeficiency virus and Leishmania belong to zymodeme MON-1. Amplification of L. infantum minicircles by the polymerase chain reaction (PCR) and digestion of the amplified product to reveal restriction fragment length polymorphisms (RFLP) has proved very useful in distinguishing between relapses and reinfections in co-infected, treated patients. We have confirmed the existence of a leishmaniasis outbreak among intravenous drug users in north-east Spain, previously detected by isoenzymatic analysis. We have documented persistence of the same strain of Leishmania in 2 treated co-infected patients throughout several years, regardless of the theoretical rapid evolution ascribed to kinetoplast deoxyribonucleic acid minicircle sequences. We suggest using this PCR-RFLP technique to detect reinfections in treated co-infected subjects.

AIDS-Related Opportunistic Infections↗
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Recombinant antigens for specific and sensitive serodiagnosis of Latin American tegumentary leishmaniasis.

The diagnostic potential of recombinant leishmanial antigens for Latin American tegumentary leishmaniasis (LATL) was examined. Two Leishmania (Viannia) peruviana recombinant proteins, T26-U2 and T26-U4, were assessed by their reactivity to detect specific anti-leishmanial antibodies. Seventy-eight individual sera from persons with LATL, 39 from those with other diseases, and 10 negative control sera were tested by Western blotting and enzyme-linked immunosorbent assay. The sensitivity of the test using T26-U2 plus T26-U4 was similar to that obtained with whole parasite extract (92%). However, the specificity obtained using both recombinant antigens (87%) was higher than that of the whole parasite extract (65%). All tests using recombinant proteins (T26-U2, T26-U2 plus T26-U4 or T26-U4) had a higher positive predictive value (89%, 92% and 98%, respectively) than the value obtained using total parasites (81%). Eleven Colombian sera were also tested, and the results indicated that T26-U2 plus T26-U4 could be used successfully in Peru and in other Latin American countries.

Animals↗
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Detection of Leishmania braziliensis in naturally infected individual sandflies by the polymerase chain reaction.

The natural infection of sandflies by Leishmania in wild-caught specimens was studied, using the polymerase chain reaction (PCR)-hybridization technique. The PCR was carried out using 2 oligonucleotides (primers 3J1 and 3J2) derived from a repetitive nuclear DNA sequence. The primers support the enzymatic amplification of a fragment of approximately 500 bp, present in the nuclear DNA of Leishmania braziliensis. The expected band was observed in 5 of 65 sandflies containing flagellates. After hybridization with a species-specific probe, we confirmed natural infection by L. braziliensis. The technique allowed the identification of Lutzomyia gomezi and Lu. panamensis as vectors of L. braziliensis in an endemic area of cutaneous leishmaniasis in Urama, Puerto Cabello district in Venezuela. As far as we are aware, this work constitutes the first report of natural infection of Lu. panamensis with L. braziliensis in the study area. We also demonstrate that PCR-hybridization is a suitable approach to establish the Leishmania-sandfly relationship and will be useful in epidemiological studies of leishmaniasis in endemic areas.

Animals↗