Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2

Keegan, Frank P.; Blum, J. J.

doi:10.1016/0166-6851(91)90175-6

Cited by 7 publications

(4 citation statements)

References 9 publications

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“…Enrichment of RPMI 1640 medium with 20% FBS resulted in an 18% improvement in microculture sensitivity compared with previous analyses in which RPMI 1640 medium containing 10% FBS was used. 18 That microaerophilia and lower O 2 :CO 2 ratios are important for amastigote-topromastigote conversion 11,[19][20][21] is supported by the success of miniculture compared with RPMI 1640 medium containing 10% FBS in a traditional culture system. 18 In addition to improved sensitivity, time-to-culture positivity was also reduced with both microculture and miniculture, which afforded a two-day incubation savings compared with traditional NNN medium culture.…”

Section: Discussionmentioning

confidence: 99%

“…11,[15][16][17][18] The higher sensitivity of microculture is thought to be related to the microaerophilic conditions and high CO 2 levels established within the system, both of which are permissive to amastigote-to-promastigote transformation. 11,[19][20][21] One limitation of microculture is that the organism, once isolated, is relatively inaccessible because of the sealed ends of the capillary tubes. We postulated that a monophasic liquid medium in 1.5-mL Eppendorf (Hamburg, Germany) tubes (miniculture) may capture the advantages of microculture and traditional culture systems.…”

Section: Introductionmentioning

confidence: 99%

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Optimization of Microculture and Evaluation of Miniculture for the Isolation of Leishmania Parasites from Cutaneous Lesions in Peru

Boggild¹,

Miranda‐Verástegui²,

Espinosa³

et al. 2008

The American Journal of Tropical Medicine and Hygiene

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Traditional culture of Leishmania parasites is labor-intensive and shows poor sensitivity. We evaluated microculture and novel miniculture methods for diagnosis of cutaneous leishmaniasis (CL). Consecutive patients who came to the Leishmaniasis Clinic, Hospital Nacional Cayetano Heredia, Lima, Peru, were enrolled. Lesion aspirates were cultured in traditional tubes containing Novy-MacNeal-Nicolle medium and in miniculture tubes (Eppendorf, Hamburg, Germany) and capillary tubes (microculture) containing RPMI 1640 medium containing 20% fetal bovine serum. The reference standard was positive results in two of four tests (smear, culture, polymerase chain reaction, or leishmanin skin test). Outcome measures were sensitivity and time to positivity. Fifty-five patients with 74 lesions were enrolled. Of 59 lesions that fulfilled reference criteria for CL, 50 were positive by microculture (sensitivity=84.7%; P=0.001), 45 by miniculture (sensitivity=76.3%; P=0.042), and 35 by traditional culture (sensitivity=59.3%). Median time to positivity was three days by microculture and miniculture and five days by traditional culture (P<0.001). Microculture and miniculture are sensitive and efficient means of diagnosing CL.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optimization of Microculture and Evaluation of Miniculture for the Isolation of Leishmania Parasites from Cutaneous Lesions in Peru

Boggild¹,

Miranda‐Verástegui²,

Espinosa³

et al. 2008

The American Journal of Tropical Medicine and Hygiene

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“…RPMI 1640 medium supplemented with 10 to 20% fetal bovine serum has been shown to support amastigote-to-promastigote conversion, though with lower sensitivity than other traditional methods (1,2,17), possibly due to the high content of dissolved oxygen in this type of culture system. Microaerophilic conditions and high CO 2 levels are permissive to amastigote-to-promastigote transformation (20,21,39), hence the success of the microculture system (2). Traditional cultures of lesion aspirates, scrapings, and biopsy samples using NNN medium, Tobie's medium, Schneider's Drosophila medium, or M199 Leishmania culture medium typically have sensitivities that range from 40 to 75%, though this is somewhat species dependent, with L. (V.) braziliensis constituting one of the more difficult species to culture (32,37).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of a Microculture Method for Isolation of Leishmania Parasites from Cutaneous Lesions of Patients in Peru

et al. 2007

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“…These results are consistent with results obtained by others. [28][29][30][31][32] High CO 2 levels may act as the trigger for amastigote-to-promastigote transformation and favor the survival of promastigotes. 33,34 It is known that CO 2 and low concentrations of O 2 have regulatory effects on glucose metabolism in promastigotes.…”

Section: Discussionmentioning

confidence: 99%

A Sensitive New Microculture Method for Diagnosis of Cutaneous Leishmaniasis

et al. 2004

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A sensitive microcapillary culture method (MCM) was developed for rapid diagnosis of cutaneous leishmaniasis (CL). The MCM is superior to the traditional culture method (TCM) as determined by the smaller inoculum size, the higher sensitivity for detection of promastigotes, and the more rapid time for emergence of promastigotes. With lesion amastigote loads from grade III to 0, the positive rates and the periods for promastigote emergence were 3-4-fold higher and faster with the MCM than with the TCM, e.g., 83-97% positive in 4-7 days versus 20-40% positive in 15-30 days (P = 0.0001). The higher Pco(2) and lower Po(2) and pH presumably encourage a rapid amastigote-to-promastigote differentiation and/or the survival or growth of the latter. This MCM has the advantage of simplicity, and may be suitable for diagnostic use and for parasite retrieval in many other endemic sites where parasites are known to be difficult to grow.

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Changes in intracellular levels of fructose 2,6-bisphosphate and several glycolytic intermediates in Leishmania major promastigotes as a function of pO2

Cited by 7 publications

References 9 publications

Optimization of Microculture and Evaluation of Miniculture for the Isolation of Leishmania Parasites from Cutaneous Lesions in Peru

Optimization of Microculture and Evaluation of Miniculture for the Isolation of Leishmania Parasites from Cutaneous Lesions in Peru

Evaluation of a Microculture Method for Isolation of Leishmania Parasites from Cutaneous Lesions of Patients in Peru

A Sensitive New Microculture Method for Diagnosis of Cutaneous Leishmaniasis

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