Appropriate transfer of molecuar technology to latin america for publica health and biomedical science

Harris, Eva; Belli, Alejandro; Agabian, Nina

doi:10.1016/0307-4412(95)00154-9

Cited by 9 publications

(10 citation statements)

References 11 publications

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“…[19][20][21][22] At the Leishmania Reference Laboratory of the Nicaraguan Ministry of Health, we have implemented the PCR for routine use because of the limitations we experienced with traditional diagnostic and characterization techniques in our field work. 10 Several reported PCR protocols were useful; 13, 19 however, we found it necessary to further simplify sample preparation methods and amplification protocols. Here we describe the adaptations developed to streamline routine use of the PCR for detection and characterization of Leishmania in endemic countries.…”

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confidence: 99%

“…The PCR is a sensitive and rapid technique, which can be adapted for use under conditions of limited resources by applying a lowcost approach. [10][11][12] A number of PCR protocols have been reported for the detection of Leishmania that cause cutaneous leishmaniasis, using either purified DNA [13][14][15][16][17][18] or clinical specimens. [19][20][21][22] At the Leishmania Reference Laboratory of the Nicaraguan Ministry of Health, we have implemented the PCR for routine use because of the limitations we experienced with traditional diagnostic and characterization techniques in our field work.…”

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confidence: 99%

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Simplified polymerase chain reaction detection of new world Leishmania in clinical specimens of cutaneous leishmaniasis.

Belli

Rodríguez²,

Avilés³

et al. 1998

Am J Trop Med Hyg

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Polymerase chain reaction (PCR)-based detection of New World Leishmania from different types of clinical specimens has been further streamlined for field use by simplifying sample preparation and modifying published protocols. A multiplex PCR reaction was developed that allows simultaneous detection of the Leishmania genus and identification of the L. braziliensis complex. For typing isolates in culture, we found that simply boiling diluted cultured strains was sufficient preparation for the PCR. We have demonstrated that Leishmania parasites can be reliably detected from boiled dermal scrapings, instead of the more invasive skin biopsies often used as PCR specimens. The PCR of dermal scrapings yielded a sensitivity of 100% and a specificity of 100%, as compared with microscopic examination. In a study population, PCR was more sensitive than classic diagnostic techniques. The PCR detection of Leishmania in biopsies and peripheral blood mononuclear cells (PBMCs) was investigated. Diluting crude extracts of skin biopsies was sufficient to eliminate sample inhibition yet maintain required sensitivity. Leishmania braziliensis was also detected by PCR in PBMCs of individuals with active cutaneous leishmaniasis. The simplifications described here significantly improve the feasibility of using the PCR in endemic countries as the primary method for detection and preliminary characterization of Leishmania in clinical specimens of cutaneous leishmaniasis.

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confidence: 99%

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confidence: 99%

Simplified polymerase chain reaction detection of new world Leishmania in clinical specimens of cutaneous leishmaniasis.

Belli

Rodríguez²,

Avilés³

et al. 1998

Am J Trop Med Hyg

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“…10 Because of the molecular specificity of PCR, detection and genetic characterization of Leishmania can be carried out simultaneously. 11,12 The extensive literature on the molecular diagnosis of ACL has provided various alternatives, from the use of primers that amplify sequences of specific Leishmania species to primers that amplify sequences common to all the species. 13,14 The objective of this study was to analyze different PCR primers that are commonly used in epidemiological studies for the detection of Leishmania, and to compare them in the diagnosis of ACL in biological samples taken from dogs and humans in an endemic region.…”

Section: Introductionmentioning

confidence: 99%

Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis

Oliveira¹,

Lonardoni²,

Teodoro³

et al. 2011

Braz J Infect Dis

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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 and methods: 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 10 3 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.

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“…This group of young, progressive researchers craved the chance to teach and apply their scientific knowledge in a way that would make a concrete difference in the lives of developing‐country residents. For 10 years, they did so through the AMB/ATT program, following Harris's initial vision, by donating their time and seeking small grants and individual donations 6–8 . Through conducting customized training workshops and simplifying technologies onsite, they taught molecular biology‐based disease diagnosis, epidemiology, and control of infectious diseases to public health researchers and educators in Nicaragua, Ecuador, Bolivia, and Guatemala.…”

Section: Background: Ssi's Genesismentioning

confidence: 99%

“…For 10 years, they did so through the AMB/ATT program, following Harris's initial vision, by donating their time and seeking small grants and individual donations. [6][7][8] Through conducting customized training workshops and simplifying technologies onsite, they taught molecular biology-based disease diagnosis, epidemiology, and control of infectious diseases to public health researchers and educators in Nicaragua, Ecuador, Bolivia, and Guatemala. They also channeled hundreds of thousands of dollars' worth of donated laboratory equipment, supplies, and reagents to Latin American researchers and institutions.…”

Section: Background: Ssi's Genesismentioning

confidence: 99%

Sustainable Transfer of Biotechnology to Developing Countries

Coloma

Harris

2008

Annals of the New York Academy of Sciences

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Poverty and social inequalities have powerfully sculpted not only the distribution of infectious and other diseases but also the course of disease in those affected. The lack of proper diagnosis and access to adequate health services only compounds the problem. In low-resource settings, the burden of disease can be reduced if the basic human and material resources exist to support the use of low-cost interventions by appropriately trained personnel. For 20 years, the Sustainable Sciences Institute has built scientific capacity in developing countries to recognize, prevent, and respond to the threats posed by disease.

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Appropriate transfer of molecuar technology to latin america for publica health and biomedical science

Cited by 9 publications

References 11 publications

Simplified polymerase chain reaction detection of new world Leishmania in clinical specimens of cutaneous leishmaniasis.

Simplified polymerase chain reaction detection of new world Leishmania in clinical specimens of cutaneous leishmaniasis.

Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis

Sustainable Transfer of Biotechnology to Developing Countries

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