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- Name:
- William Crafts
- Affiliation:
- Michigan Department of Community Health
- City_State:
- Lansing, Michigan
- Country:
- USA
- Email:
- [email protected]
- Date:
- 2/22/00
- Time:
- 12:30:45 PM
HPLC
We have been performing HPLC since 1989
We currently use Waters HPLC system both UV and FL
We analyze approx. 1400 specimens per year
Most commons species are tuberculosis, avium, gordonae, fortuitum, Nocardia sp.
HPLC is used in addition to Gen-Probe and biochemicals for final ID.
Comments
We are currently having problems with the sensitivity of our HPLC-FL broth method as it pertains to 7H9 and Bactec 12B broth. Could someone E-mail or Fax me their current procedure for HPLC-FL or HPLC-UV broth method. We do not seem to achieve any better sensitivity when comparing UV to FL so we must be doing something wrong. I would be very interested in the procedure Ken Jost uses for his lab in Texas. Can anyone help, thanks
- Name:
- Jill Clemmer
- Affiliation:
- Massachusetts State Laboratory
- City_State:
- Jamaica Plain, MA
- Country:
- USA
- Email:
- [email protected]
- Date:
- 12/28/99
- Time:
- 8:56:34 AM
HPLC
HPLC for mycolic acids using fluorescence detection is being set-up in our lab (1999).
The system is Hewlett Packard series 1100, except for the detector which is Jasco FP-920.
Chemstation software is used for instrument control, data collection and reporting.
How the HPLC method will be used has not been determined, yet. It is not the primary
ID tool at this lab.
Comments
- Name:
- test
- Affiliation:
- City_State:
- Country:
- USA
- Email:
- Date:
- 12/2/99
- Time:
- 11:17:03 AM
HPLC
This is a test
Comments
- Name:
- Joe Aguilera
- Affiliation:
- South Texas Hospital-TDH
- City_State:
- Harlingen,Tx
- Country:
- USA
- Email:
- [email protected]
- Date:
- 10/2/99
- Time:
- 4:08:39 PM
HPLC
For now I can just tell you that we want to start doing Myclic acids profiles
for AFB Identification.
Comments
We are a Clinical Lab with a very busy Mycobacteriology seccion: We proscess all
the specimens from the Mexican State of Tamaulipas as well as the local specimens from the
Public Health Region-11.We are looking at the possibility of reducing costs and improve
our turn-around time for AFB ID.We are hopping to work very closely with Ken Jost from the
Texas Department of Health, noe the new chairman of the steering committee.
previous guest book entries
Name: Kent Molin <[email protected]>
Affiliation: Sahlgren's University Hospital
SWEDEN
Currently Using HPLC (Y or N): Y
HPLC Comments: We have been using HPLC-analysis of mycolic acids since May 1997,
using the CDC Manual. The method is used when other tests, e.g. gene-probe kits and
biochemical methods, fails to give identification. It is also sometimes used to confirm
results of other methods. We also use a GC method for fatty acid analysis. The most common
species identified by HPLC are M. malmoense, M. marinum, M. chelonae/abscessus and M.
avium complex. Altogether, 17 different species or complexes have been identified. Some of
the The method works pretty well, and is a fine complement to the other methods. We even
discovered species that had not been identified by this lab before! The results sometimes
are confusing though. There are plans for setting up a DNA sequencing method for
mycobacteria. If this is done, a study will probably be performed to investigate if HPLC
and other methods will be needed any longer.
Additional Comments:
- Monday, 27 Sep 1999 04:20:35 -0500
Name: Mike Garvey <[email protected]>
Alton, IL USA
Currently Using HPLC (Y or N): Y
HPLC Comments:
Additional Comments:
- Thursday, May 27, 1999 at 21:10:33 (MDT
Name: ruth ann spence <[email protected]>
Affiliation: tn. dept of health
nashville, tn USA
Currently Using HPLC (Y or N): y
Maker of the HPLC System: beckman
Type of Detection (UV or FL): uv
Number of Specimens Analyzed per Year by HPLC: ?
HPLC Comments:
The process has been used on a regular basis for three years
Other than the regular ones, we get interjectum quite often and also the ncp202-206.
The protocol is that if the organism is not in an open file(those that been identified
with a consistency verified by cdc} it is backed up with bios. Also if the organism is an
odd one not normally encountered, it is sent to cdc to verify and supplement the library.
Additional Comments:
- Saturday, February 06, 1999 at 19:17:08 (MST
Name: Louise Thibert <[email protected]>
Affiliation: Laboratoire de sant� publique du Qu�bec (Pub Hlth Lab)
Ste-Anne-de-Bellevue, QC Canada
Currently Using HPLC (Y or N): Y
Maker of the HPLC System: HP and Waters
Type of Detection (UV or FL): UV
Number of Specimens Analyzed per Year by HPLC: � 2000
HPLC Comments:
We have been using HPLC as our primary routine identification method since January 1991.
It allows us the final identification of over 90% of all isolates received, and 2/3 are
broth medium cultures (We do not receive any clinical specimens only cultures). For a very
small % of isolates, we add a few selected tests. We are in the process of setting up the
FL detection. The most common species are M. tuberculosis, M. avium complex, M. gordonae
(!!), M. fortuitum complex. HPLC is a great method for the detection and identification of
fastidious, rare or atypical mycobacterial species such as M. genavense, M. haemophilum,
M. shimoidei, M. celatum, M. triplex..... Fo us, another very important use for HPLC is
the identification to the genus, and sometimes to the species, of all Nocardia,
Rhodococcus, Gordonia and Tsukamurella isolates for which we have also eliminated most
biochemical tests.
Additional Comments:
- Thursday, January 07, 1999 at 11:05:09 (MST
Name: Enrico Tortoli <[email protected]>
Affiliation: Careggi Hospital
Florence, Italy
Currently Using HPLC (Y or N): Yes
Maker of the HPLC System: Beckman
Type of Detection (UV or FL): UV
Number of Specimens Analyzed per Year by HPLC: 500
HPLC Comments:
We started using HPLC in 1993. The species we most frequemtly encounter are: M.
tuberculosis, M. xenopi, MAC, M. fortuitum, M. chelonae, M. terrae
We identify with HPLC, in combination with biochemical tests, all the species for which
probes are not available
Additional Comments:
- Thursday, December 17, 1998 at 11:38:59 (MST
Name: Dave Bankert <[email protected]>
Affiliation: York Hospital
York, Pa USA
Currently Using HPLC (Y or N): Y
Maker of the HPLC System: HP 1100 series
Type of Detection (UV or FL): FL
Number of Specimens Analyzed per Year by HPLC: ?
HPLC Comments:
We started using HPLC last December. We're presently working through the strengths and
weakness in the system. We will be submitting a comparison study on the ability of the
MIDI software to ID mycobacteria. We hope to eliminate DNA probe for MAI and Mtb. Our most
frequent isolate is MAI.
Additional Comments:
- Wednesday, November 11, 1998 at 19:06:11 (MST
Name: Linda Guthertz <[email protected]>
Affiliation: CDHS-Microbial Diseases Lab
Berkeley , CA USA
Currently Using HPLC (Y or N): y
Maker of the HPLC System: Waters, Hewlett-Packard
Type of Detection (UV or FL): both
Number of Specimens Analyzed per Year by HPLC: lots
HPLC Comments:
Additional Comments:
- Thursday, October 22, 1998 at 07:38:51 (MDT
Name: gary cage <[email protected]>
Affiliation: arizona state laboratory
phoenix, az USA
Currently Using HPLC (Y or N): yes
Maker of the HPLC System: waters mix and match
Type of Detection (UV or FL): both
Number of Specimens Analyzed per Year by HPLC: 5200
HPLC Comments:
we've been using hplc since about 1990 using uv detection. we started playing around with
fluorescent detection 1995. For the past year or so, we've been using HPLC-FL for
identification from direct, smear-positive specimens, based on Ken Jost's work. Our most
frequent species are TB, MAC and simiae. We've identified about 40 different species using
this method. We've used HPLC EXCLUSIVELY for CAP unknowns since 1991, and have not missed
one since, so you know this method has to be a good one (CAP finially got around to
listing chromatography as an ID "Method").
Additional Comments:
Despite fine tuning problems with the HPLC method, the overall usefullness, I feel, is
great. Unfortunately, it does seem to be under-represented in the mainstream TB
literature, especially for the clinical laboratory. That's why I feel that the group and
its membership is extremely important in getting the word out! I urge those who use HPLC
publish their findings/experiences to raise awareness in the TB community.
- Monday, October 19, 1998 at 01:29:58 (MDT
Name: James R. (Jim) Meldrum <[email protected]>
Affiliation: State Health Lab/UNR Med School
Reno, Nv USA
Currently Using HPLC (Y or N): Y
Maker of the HPLC System: Beckman System Gold
Type of Detection (UV or FL): UV
Number of Specimens Analyzed per Year by HPLC: 500
HPLC Comments:
Our HPLC has been in use for about 6 months as a primary identification tool for
Mycobacteria isolated from sputum or reference cultures submitted to the laboratory.
Analysis is performed on cultures from solid media and liquid media (Bactec). We typically
isolate MTB, avium, intracellulare, scrof., gordonae, fort./perig., marinum, and a few
others.
Additional Comments:
- Thursday, October 15, 1998 at 13:37:36 (MDT
Name: SEMPERE ALCOCER MARCO ANTONIO <[email protected]>
Affiliation: PHD IN COSTA DEL SOL HOSPITAL
MARBELLA (MALAGA), SPAIN
Currently Using HPLC (Y or N): YES
Maker of the HPLC System: WATERS
Type of Detection (UV or FL): both
Number of Specimens Analyzed per Year by HPLC: 200
HPLC Comments:
since 1997
M.tuberculosis, M.fortuitum and M.chelonae
Yes is the primary identification tool
I use the patern recognition of Infometrix with Waters HPLC
Additional Comments:
- Thursday, August 06, 1998 at 10:58:23 (MDT
Name: Cho, Myung-Lae <[email protected]>
Currently Using HPLC (Y or N): N
HPLC Comments:
Additional Comments:
- Tuesday, July 28, 1998 at 23:53:42 (MDT
Revised: 30 Jun 2000 14:42:41 -0500.