Evaluation of a Microculture Method for Isolation of Leishmania Parasites from Cutaneous Lesions of Patients in Peru
Traditional culture of Leishmania spp. is labor intensive and has poor sensitivity. We evaluated a microculture method for the diagnosis of cutaneous leishmaniasis in consecutive patients presenting to the Leishmaniasis Clinic at the Instituto de Medicina Tropical Alexander von Humboldt, Peru, for e...
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| Veröffentlicht in: | Journal of Clinical Microbiology 2007-11, Vol.45 (11), p.3680-3684 |
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| creator | Boggild, Andrea K Miranda-Verastegui, Cesar Espinosa, Diego Arevalo, Jorge Adaui, Vanessa Tulliano, Gianfranco Llanos-Cuentas, Alejandro Low, Donald E |
| description | Traditional culture of Leishmania spp. is labor intensive and has poor sensitivity. We evaluated a microculture method for the diagnosis of cutaneous leishmaniasis in consecutive patients presenting to the Leishmaniasis Clinic at the Instituto de Medicina Tropical Alexander von Humboldt, Peru, for evaluation of skin lesions. Lesion aspirates were cultured in duplicate and parallel in traditional culture tubes containing modified Novy-MacNeal-Nicolle (NNN) medium or Roswell Park Memorial Institute medium 1640 with 10% fetal bovine serum (10% RPMI) and in 70-μl capillary tubes containing a mixture of lesion aspirate and 10% RPMI. For sensitivity analysis, the consensus standard was considered to be a positive result in any two of the following four tests: Giemsa-stained lesion smear, culture, kinetoplast DNA PCR, or leishmanin skin test. The outcome measures were sensitivity and time to culture positivity. Forty-five patients with 62 skin lesions were enrolled in the study, of which 53 lesions fulfilled the consensus criteria for a final diagnosis of cutaneous leishmaniasis. Of these 53 lesions, 39 were culture positive: 38 in capillary tubes, 29 in traditional culture tubes with modified NNN medium, and 19 in traditional culture tubes with 10% RPMI medium. The sensitivity of microculture was 71.7%, versus 54.7% for traditional culture with NNN (P, 0.038) and 35.8% with 10% RPMI (P, |
| doi_str_mv | 10.1128/JCM.01286-07 |
| format | Article |
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We evaluated a microculture method for the diagnosis of cutaneous leishmaniasis in consecutive patients presenting to the Leishmaniasis Clinic at the Instituto de Medicina Tropical Alexander von Humboldt, Peru, for evaluation of skin lesions. Lesion aspirates were cultured in duplicate and parallel in traditional culture tubes containing modified Novy-MacNeal-Nicolle (NNN) medium or Roswell Park Memorial Institute medium 1640 with 10% fetal bovine serum (10% RPMI) and in 70-μl capillary tubes containing a mixture of lesion aspirate and 10% RPMI. For sensitivity analysis, the consensus standard was considered to be a positive result in any two of the following four tests: Giemsa-stained lesion smear, culture, kinetoplast DNA PCR, or leishmanin skin test. The outcome measures were sensitivity and time to culture positivity. Forty-five patients with 62 skin lesions were enrolled in the study, of which 53 lesions fulfilled the consensus criteria for a final diagnosis of cutaneous leishmaniasis. Of these 53 lesions, 39 were culture positive: 38 in capillary tubes, 29 in traditional culture tubes with modified NNN medium, and 19 in traditional culture tubes with 10% RPMI medium. The sensitivity of microculture was 71.7%, versus 54.7% for traditional culture with NNN (P, 0.038) and 35.8% with 10% RPMI (P, <0.001). The mean times to culture positivity were 4.2 days by microculture, 5.2 days in NNN, and 6 days in 10% RPMI (P, 0.009). We have demonstrated that microculture is a more sensitive and time-efficient means of isolating Leishmania parasites from cutaneous lesions than traditional culture.</description><identifier>ISSN: 0095-1137</identifier><identifier>EISSN: 1098-660X</identifier><identifier>EISSN: 1098-5530</identifier><identifier>DOI: 10.1128/JCM.01286-07</identifier><identifier>PMID: 17881557</identifier><identifier>CODEN: JCMIDW</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Biological and medical sciences ; Child ; Child, Preschool ; DNA, Protozoan - analysis ; Female ; Human protozoal diseases ; Humans ; Infant ; Infectious diseases ; Leishmania ; Leishmania - isolation & purification ; Leishmaniasis, Cutaneous - diagnosis ; Leshmaniasis ; Male ; Medical sciences ; Middle Aged ; Parasitic diseases ; Parasitology ; Polymerase Chain Reaction ; Protozoal diseases ; Sensitivity and Specificity ; Skin - parasitology</subject><ispartof>Journal of Clinical Microbiology, 2007-11, Vol.45 (11), p.3680-3684</ispartof><rights>2008 INIST-CNRS</rights><rights>Copyright © 2007, American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c494t-149a42fd726186fd8fdd82e3eb5f269e37d029dd1aad0e49e3946a9cc8ada5993</citedby><cites>FETCH-LOGICAL-c494t-149a42fd726186fd8fdd82e3eb5f269e37d029dd1aad0e49e3946a9cc8ada5993</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2168488/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2168488/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,3189,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19372541$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17881557$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Boggild, Andrea K</creatorcontrib><creatorcontrib>Miranda-Verastegui, Cesar</creatorcontrib><creatorcontrib>Espinosa, Diego</creatorcontrib><creatorcontrib>Arevalo, Jorge</creatorcontrib><creatorcontrib>Adaui, Vanessa</creatorcontrib><creatorcontrib>Tulliano, Gianfranco</creatorcontrib><creatorcontrib>Llanos-Cuentas, Alejandro</creatorcontrib><creatorcontrib>Low, Donald E</creatorcontrib><title>Evaluation of a Microculture Method for Isolation of Leishmania Parasites from Cutaneous Lesions of Patients in Peru</title><title>Journal of Clinical Microbiology</title><addtitle>J Clin Microbiol</addtitle><description>Traditional culture of Leishmania spp. is labor intensive and has poor sensitivity. We evaluated a microculture method for the diagnosis of cutaneous leishmaniasis in consecutive patients presenting to the Leishmaniasis Clinic at the Instituto de Medicina Tropical Alexander von Humboldt, Peru, for evaluation of skin lesions. Lesion aspirates were cultured in duplicate and parallel in traditional culture tubes containing modified Novy-MacNeal-Nicolle (NNN) medium or Roswell Park Memorial Institute medium 1640 with 10% fetal bovine serum (10% RPMI) and in 70-μl capillary tubes containing a mixture of lesion aspirate and 10% RPMI. For sensitivity analysis, the consensus standard was considered to be a positive result in any two of the following four tests: Giemsa-stained lesion smear, culture, kinetoplast DNA PCR, or leishmanin skin test. The outcome measures were sensitivity and time to culture positivity. Forty-five patients with 62 skin lesions were enrolled in the study, of which 53 lesions fulfilled the consensus criteria for a final diagnosis of cutaneous leishmaniasis. Of these 53 lesions, 39 were culture positive: 38 in capillary tubes, 29 in traditional culture tubes with modified NNN medium, and 19 in traditional culture tubes with 10% RPMI medium. The sensitivity of microculture was 71.7%, versus 54.7% for traditional culture with NNN (P, 0.038) and 35.8% with 10% RPMI (P, <0.001). The mean times to culture positivity were 4.2 days by microculture, 5.2 days in NNN, and 6 days in 10% RPMI (P, 0.009). We have demonstrated that microculture is a more sensitive and time-efficient means of isolating Leishmania parasites from cutaneous lesions than traditional culture.</description><subject>Adolescent</subject><subject>Adult</subject><subject>Aged</subject><subject>Aged, 80 and over</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Child</subject><subject>Child, Preschool</subject><subject>DNA, Protozoan - analysis</subject><subject>Female</subject><subject>Human protozoal diseases</subject><subject>Humans</subject><subject>Infant</subject><subject>Infectious diseases</subject><subject>Leishmania</subject><subject>Leishmania - isolation & purification</subject><subject>Leishmaniasis, Cutaneous - diagnosis</subject><subject>Leshmaniasis</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Middle Aged</subject><subject>Parasitic diseases</subject><subject>Parasitology</subject><subject>Polymerase Chain Reaction</subject><subject>Protozoal diseases</subject><subject>Sensitivity and Specificity</subject><subject>Skin - parasitology</subject><issn>0095-1137</issn><issn>1098-660X</issn><issn>1098-5530</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0c1vFCEUAHBiNHZbvXlWYqInpwIDM3AxMZuqNbtxE23ijbzysUszM7Qw08b_XtbZtHryBIEfj_eB0AtKTill8v3X5fqUlE1TkfYRWlCiZNU05OdjtCBEiYrSuj1CxzlfEUI5F-IpOqKtlFSIdoHGs1voJhhDHHD0GPA6mBTN1I1Tcnjtxl202MeEz3Ps7tnKhbzrYQiAN5Agh9Fl7FPs8XIaYXBxysXkovOeb8pDN4wZhwFvXJqeoSceuuyeH9YTdPHp7MfyS7X69vl8-XFVGa74WFGugDNvW9ZQ2XgrvbWSudpdCs8a5erWEqaspQCWOF4OFG9AGSPBglCqPkEf5rjX02XvrCk5JOj0dQo9pF86QtD_3gxhp7fxVjPaSC5lCfD2ECDFm8nlUfchG9d1c426qFaxWvwXMiKYEmwP382wdDnn5Px9NpTo_Tx1maf-M09N2sJf_l3BAz4MsIA3BwDZQOcTDCbkB6fqlglOi3s9u13Y7u5Cchpyr69Mr7ko_-q6kaSgVzPyEDVsUwl08Z0RWhMimSgdqX8DPEO_Wg</recordid><startdate>20071101</startdate><enddate>20071101</enddate><creator>Boggild, Andrea K</creator><creator>Miranda-Verastegui, Cesar</creator><creator>Espinosa, Diego</creator><creator>Arevalo, Jorge</creator><creator>Adaui, Vanessa</creator><creator>Tulliano, Gianfranco</creator><creator>Llanos-Cuentas, Alejandro</creator><creator>Low, Donald E</creator><general>American Society for Microbiology</general><general>American Society for Microbiology (ASM)</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>M7N</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20071101</creationdate><title>Evaluation of a Microculture Method for Isolation of Leishmania Parasites from Cutaneous Lesions of Patients in Peru</title><author>Boggild, Andrea K ; 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We evaluated a microculture method for the diagnosis of cutaneous leishmaniasis in consecutive patients presenting to the Leishmaniasis Clinic at the Instituto de Medicina Tropical Alexander von Humboldt, Peru, for evaluation of skin lesions. Lesion aspirates were cultured in duplicate and parallel in traditional culture tubes containing modified Novy-MacNeal-Nicolle (NNN) medium or Roswell Park Memorial Institute medium 1640 with 10% fetal bovine serum (10% RPMI) and in 70-μl capillary tubes containing a mixture of lesion aspirate and 10% RPMI. For sensitivity analysis, the consensus standard was considered to be a positive result in any two of the following four tests: Giemsa-stained lesion smear, culture, kinetoplast DNA PCR, or leishmanin skin test. The outcome measures were sensitivity and time to culture positivity. Forty-five patients with 62 skin lesions were enrolled in the study, of which 53 lesions fulfilled the consensus criteria for a final diagnosis of cutaneous leishmaniasis. Of these 53 lesions, 39 were culture positive: 38 in capillary tubes, 29 in traditional culture tubes with modified NNN medium, and 19 in traditional culture tubes with 10% RPMI medium. The sensitivity of microculture was 71.7%, versus 54.7% for traditional culture with NNN (P, 0.038) and 35.8% with 10% RPMI (P, <0.001). The mean times to culture positivity were 4.2 days by microculture, 5.2 days in NNN, and 6 days in 10% RPMI (P, 0.009). We have demonstrated that microculture is a more sensitive and time-efficient means of isolating Leishmania parasites from cutaneous lesions than traditional culture.</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>17881557</pmid><doi>10.1128/JCM.01286-07</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Adolescent Adult Aged Aged, 80 and over Animals Biological and medical sciences Child Child, Preschool DNA, Protozoan - analysis Female Human protozoal diseases Humans Infant Infectious diseases Leishmania Leishmania - isolation & purification Leishmaniasis, Cutaneous - diagnosis Leshmaniasis Male Medical sciences Middle Aged Parasitic diseases Parasitology Polymerase Chain Reaction Protozoal diseases Sensitivity and Specificity Skin - parasitology |
| title | Evaluation of a Microculture Method for Isolation of Leishmania Parasites from Cutaneous Lesions of Patients in Peru |
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