Rapid identification of non-sporing anaerobes using nuclear magnetic resonance spectroscopy and an identification strategy.
OBJECTIVE
The non-sporing anaerobes cause a wide spectrum of infections. They are difficult to culture and their identification is tedious and time-consuming. Rapid identification of anaerobes is highly desirable. Towards this end, the potential of nuclear magnetic resonance (NMR) spectroscopy for providing a fingerprint within the proton spectrum of six genera belonging to anaerobes reflecting their characteristic metabolites has been investigated.


METHODS
NMR analysis was carried out using Mercury plus Varian 300 MHz (7.05 T) NMR spectrophotometer on six different anaerobes. These included Bacteroides fragilis, Prevotella melaninogenica, Prevotella denticola, Fusobacterium necrophorum, Peptococcus niger and Peptostreptococcus spp. After the NMR analysis (256/512 scans), the different peaks were noted. The eight pus specimens, which yielded pure culture of anaerobe, also were analysed similarly.


RESULTS
The major resonances of multiplex of amino acids/lipid at 0.9 ppm along with lactate/lipid at 1.3 ppm, acetate at 1.92 ppm and multiplex of lysine at 3.0 ppm remained constant to label the organism as an anaerobe. There was a difference found in the MR spectra of different genera and species. A simple algorithm was developed for the identification of the six different anaerobes studied. The MR spectra of the pure culture of the organism matched the MR spectra of pus from which the organism was isolated.


CONCLUSIONS
MR-based identification was of value in the identification of anaerobes. However, a larger database of the peaks produced by anaerobes needs to be created for identification of all genera and species. It could then have the potential of diagnosing an anaerobic infection in vivo and thus expedite management of deep-seated abscesses.
Menon
S
S
Department of Microbiology, Grant Medical College and Sir J.J. Hospital, Mumbai - 400 008, Maharashatra, India. saralagopalmenon@yahoo.co.in
Bharadwaj
R
R
Chowdhary
A S
AS
Kaundinya
D V
DV
Palande
D A
DA
D019342:Acetic Acid / Q000032:analysis; D000465:Algorithms; D000596:Amino Acids / Q000032:analysis; D001421:Bacteria, Anaerobic / Q000737:chemistry* / Q000302:isolation & purification*; D001424:Bacterial Infections / Q000175:diagnosis*; D003937:Diagnosis, Differential; D006801:Humans; D019344:Lactic Acid / Q000032:analysis; D008055:Lipids / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D013492:Suppuration / Q000382:microbiology
D000596:Amino Acids; D008055:Lipids; D019344:Lactic Acid; D019342:Acetic Acid