Work

Universal Genotyping in Tuberculosis Control Program, New York City, 2001–2003

Year: 2006

Type: article

Abstract: providers of new requirements; 3) modifying laboratory submission forms; 4) establishing a specimen transport system; and 5) tracking and reviewing all submissions.In addition, protocols were developed for surveillance of genotype results and false-positive culture investigations, existing patient interview forms were modified, new databases were created, and program staff were informed through special trainings and newsletters.The New York City Department of Health and Mental Hygiene Institutional Review Board and the associate director for Science of the National Center for HIV, STD, and TB Prevention, Centers for Disease Control and Prevention, reviewed the protocols, procedures, and modified data forms and determined that the genotyping service was not human subjects research since it would become a routine program activity. Laboratory ProceduresAn additional full-time staff person was hired by the Bureau of Tuberculosis Control to coordinate genotyping services.Spoligotyping and RFLP, respectively, were performed by the New York State Department of Health's Wadsworth Center in Albany, New York, and the Public Health Research Institute in Newark, New Jersey.This combination of genotyping methods is sensitive and specific for determining matching genotypes (20-24).Isolates of Mycobacterium tuberculosis complex submitted to the public health laboratory from clinical laboratories were received on solid or liquid media and were stored at 4°C.Liquid media were prepared (10% glycerol in Dubos Davis broth with Tween and albumin), 1 mL of liquid culture was injected and incubated for 3 days at 37°C and checked visually for growth.Four freeze vials (1 for spoligotyping and 3 for archiving) and 1 Lowenstein-Jensen slant were injected.Mycobacteria in the tubes for spoligotyping were heat-killed at 80°C for 1 hour and mailed in biohazard containers to the Wadsworth Center.Once appropriate growth was obtained on the Lowenstein-Jensen slants, they were sent in a biohazard container to the Public Health Research Institute for IS6110 RFLP analysis.Packages were mailed on a weekly or biweekly basis, depending on the number of isolates received.Spoligotype analysis was performed at the Wadsworth Center and given descriptive nomenclature according to a standard method (25-27).DNA analysis based on IS6110 Southern blot hybridization was performed at the Public Health Research Institute with previously described methods (28,29).To ensure good communication, a working group of all partners in the genotyping service was formed.Regular telephone conferences were conducted to address issues such as quality and shipping of isolates and submission time for genotyping.

Source: Emerging infectious diseases

DOI: 10.3201/eid1205.050446