PMID- 8345507
OWN - NLM
STAT- MEDLINE
DCOM- 19930907
LR  - 20131121
IS  - 0022-2615 (Print)
IS  - 0022-2615 (Linking)
VI  - 39
IP  - 2
DP  - 1993 Aug
TI  - Detection of Staphylococcus aureus with biotinylated monoclonal antibodies 
      directed against staphylococcal TNase complexed to avidin-peroxidase in a rapid 
      sandwich enzyme-linked immunofiltration assay (sELIFA).
PG  - 128-34
AB  - For rapid identification of Staphylococcus aureus, a monoclonal antibody 
      (MAb)-biotin-avidin-peroxidase complex, directed against the S. aureus 
      thermostable nuclease (TNase), was formed and used in a rapid three-step sandwich 
      enzyme-linked immunofiltration assay (sELIFA) and a three-step sandwich 
      enzyme-linked immunosorbent assay (sELISA). The MAb-peroxidase complex was formed 
      by incubating the biotinylated MAbs with a streptavidin-peroxidase conjugate and 
      the complex was purified by gel permeation chromatography. When compared with a 
      four-step MAb-based sELISA described previously, this complex permitted one 
      reagent step to be omitted in a three-step sELISA, and the test time was 
      significantly reduced. The test sensitivity was slightly reduced in the 
      three-step ELISA (detection limit 1.0-2.0 ng of TNase/ml) when compared to the 
      four-step sELISA (detection limit 0.5-1.0 ng of TNase/ml). The sELIFA method was 
      based on the filtration of bacterial culture supernates through nitrocellulose 
      membrane disks pre-spotted with a MAb directed against the S. aureus TNase, 
      followed by detection with the MAb-peroxidase complex (three-step sELIFA). A 
      detection limit of 0.5-2.0 ng of TNase/ml was achieved with the three-step 
      sELIFA, depending on the filtrate volume of culture supernates. The total test 
      time was 10-15 min when pre-spotted and blocked membranes were used. A total of 
      85 bacterial strains was tested in the sELIFA. All the 28 S. aureus strains 
      showed positive results, but none of the 57 non-S. aureus strains did so, 
      although some of these produced thermostable nuclease activity.(ABSTRACT 
      TRUNCATED AT 250 WORDS)
FAU - Brakstad, O G
AU  - Brakstad OG
AD  - Applied Chemistry Division, SINTEF, Trondheim, Norway.
FAU - Maeland, J A
AU  - Maeland JA
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Med Microbiol
JT  - Journal of medical microbiology
JID - 0224131
RN  - 0 (Antibodies, Monoclonal)
RN  - 1405-69-2 (Avidin)
RN  - 6SO6U10H04 (Biotin)
RN  - EC 3.1.31.1 (Micrococcal Nuclease)
SB  - IM
MH  - Antibodies, Monoclonal/*immunology
MH  - Avidin
MH  - Bacteremia/diagnosis/microbiology
MH  - Biotin
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Humans
MH  - Immunoenzyme Techniques
MH  - Micrococcal Nuclease/*immunology
MH  - Sensitivity and Specificity
MH  - Staphylococcal Infections/diagnosis/microbiology
MH  - Staphylococcus aureus/enzymology/immunology/*isolation & purification
EDAT- 1993/08/01 00:00
MHDA- 1993/08/01 00:01
CRDT- 1993/08/01 00:00
PHST- 1993/08/01 00:00 [pubmed]
PHST- 1993/08/01 00:01 [medline]
PHST- 1993/08/01 00:00 [entrez]
AID - 10.1099/00222615-39-2-128 [doi]
PST - ppublish
SO  - J Med Microbiol. 1993 Aug;39(2):128-34. doi: 10.1099/00222615-39-2-128.