Introduction of Methyl-α-D-Glucopyranoside Test
Methyl-α-D-Glucopyranoside Test short form is MGP test. MGP test is effective for species identification of Enterococcus. Enterococcus gallinarum and Enterococcus casseliﬂavus are MGP test positive while Enterococcus faecalis and Enterococcus faecium are negative. Vancomycin-resistant Enterococcus faecium (VREF) is a serious nosocomial problem that is difficult to differentiate from Enterococcus gallinarum and Enterococcus casseliﬂavus, while not important for purposes of control of nosocomial infections, show intrinsic resistance to vancomycin due to the vanC gene. Another side, E. gallinarum, and E. casseliﬂavus are difﬁcult to differentiate from VREF by conventional biochemical tests since they are phenotypically closely related. Therefore, the MGP test is useful in preventing the misidentiﬁcation of vancomycin-resistant E. gallinarum as VREF for laboratories that cannot perform the rapid tube motility test. It is an alternative to the motility test. Typically, E. gallinarum and E. casseliﬂavus can be differentiated from E. faecium based on motility: E. gallinarum and E. casseliﬂavus are motile, while E. faecium is non-motile.
Principle of Methyl-α-D-Glucopyranoside Test
MGP broth is useful to differentiate enterococci based on the ability to acidify the carbohydrate MGP. Phenol red indicator present in a medium that helps to identify the organisms on the basis of color development i.e. development of yellow color, MGP test positive whereas red or orange negative.
Requirements for Methyl-α-D-Glucopyranoside Test
- Organisms should be tested having one of the following features-
- Test any GPC that is catalase-negative and PYR positive and grows on plates with 6 µg of vancomycin per ml or demonstrates resistance to vancomycin by antimicrobial susceptibility testing but shows susceptibility to ampicillin. Test any isolate that is identiﬁed as Enterococcus faecium by a commercial kit system but is ampicillin susceptible.
- Test any nonmotile enterococcus that demonstrates resistance to vancomycin with a MIC less than 32 µg/ml but is ampicillin susceptible.
- MGP test can be performed from any blood agar plate or any plate medium with enterococcal growth, even including media with bile-esculin or azide.
- MGP brothComposition of MGP brothPancreatic digest of casein :10.0 gSodium chloride :5.0 gMGP :10.0 gPhenol red :18.0 mgDistilled or deionized water: 1000 ml
- Inoculating needle
- Control organisms
- Positive Control(PC) E. gallinarum ATCC 49573
- Negative Control (NC) E. faecalis ATCC 29212
Procedure of MGP Test
- With the help of inoculating needle, Inoculate the MGP broth by lightly touching a single, isolated colony.
- Do not use a heavy inoculum.
- Incubate aerobically at 35°C for 24 hours.
- Observe for development of yellow color change in the medium.
Result interpretation of Methyl-α-D-Glucopyranoside Test
MGP test Positive: Development of a yellow color acidiﬁcation of MGP
MGP test Negative: Red or orange color
E. gallinarum ATCC 49573: MGP test Positive
E. faecalis ATCC 29212: MGP Negative
Limitations of Methyl-α-D-Glucopyranoside Test
- A heavy inoculum may give false-positive results.
- Longer incubation times may result in false-positive reactions.
- The test has reported sensitivity of 98-100% and a speciﬁcity of 95%
- Enterococcus vagococcus gives a positive result.
- Cowan and Steel’s, manual for the identification of medical bacteria
- Lynne S. Garcia, Clinical Microbiology Procedures Handbook