Looking for LGV in San Francisco using Real-Time PCR San Francisco Department Of Public Health.

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Presentation transcript:

Looking for LGV in San Francisco using Real-Time PCR San Francisco Department Of Public Health

Lymphogranuloma Venereum (LGV) Serovar “L” Chlamydia Tropism for lymph nodes; replicates in mononuclear phagocytes Lymphadenitis, abscesses, buboes, proctocolitis Can result in serious systemic infection, death

L Serovar : a 36 base-pair deletion of the pmpH gene (table shown from Stothard et.al. 2003)

Real-Time PCR for LGV (using the method of Morre et.al with slight modifications for our equipment and specimen collection ) Validation: Real-Time PCR compared to DNA-sequencing: Tested: Clinical specimens: -18 L-serovar -10 Non-L All detectable by PCR; (100%se / 100%sp) All confirmed by Sequencing Sensitivity of 0.01 Inclusion-forming units

Process Specimen Taken At Clinic Tested For CT By SDA or TMA Nucleic Acid Extraction / DNA purification Real-Time PCR (method of Morre et al (2005) Rectal swab Purified DNA

Technical notes MagNA Pure, Automated nucleic acid extraction -necessary even for specimens already collected in lysis buffer for TMA -”DNA Kit III (Bacteria and Fungi)” performed better than “Total Nucleic Acid Kit” LightCycler Real-Time instrument; Time to results for 32 specimens: 5-6 hours

Overall 1569 rectal-positive Chlamydia specimens tested ( ) 44 (2.8%) LGV + Type L2b, by sequencing (omp1 gene) (only sequenced 10 of them) These 1569 were found through routine screening for CT/GC Not selected on basis of symptomology Very briefly used on specimens suspected, by clinician to be LGV+ 17 specimens 4 (23.5%) LGV+

Performance issues Prevalence when SDA was Chlamydia screening test: 3.0% Prevalence when TMA was Chlamydia screening test: 2.7% Issues: (1) TMA is more sensitive than PCR; there are probably specimens that are slipping through the cracks. (TMA+ LGV pcr-negative that really were LGV) (2) There is no internal control or quantitation standard

What does all this mean ? Not much, maybe: Schacter & Moncada, 2005, STD Vol 32., No. 6: -LGV has probably long been here -Found several cases of LGV in SF in 1981 (also were L2b) -not associated with anything more severe than proctitis

What now ? Scaling back since early 2007 Are there treatment questions to be answered ? How does it get from rectum to rectum ?

Looking for Non-Rectal LGV Genital Lesions 191 lesions: -negative for HSV -negative for Syph. Zero positive for LGV Urine 244 Chlamydia- positive urines Zero positive for LGV

Thank You for your attention and time Jeffrey Klausner, Director, STD Control and Prevention, SFDPH Sally Liska, Director, SFDPH Laboratory John Papp, CDC Angelique O’Connor, CDC Charlotte Kent, SFDPH (now with CDC) Lina Castro, SFDPH Laboratory Alyssa Ren, CDC/APHL Fellowship Program Acknowledgements: