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Gram stain (Gram staining or Gram's method), is a method of staining used to classify bacterial species into two large groups: gram-positive bacteria and gram-negative bacteria. It may also be used to diagnose a fungal infection. [1] The name comes from the Danish bacteriologist Hans Christian Gram, who developed the technique in 1884. [2] Gram ...
In bacteriology, gram-positive bacteria are bacteria that give a positive result in the Gram stain test, which is traditionally used to quickly classify bacteria into two broad categories according to their type of cell wall. The Gram stain is used by microbiologists to place bacteria into two main categories, Gram-positive (+) and Gram ...
A Gram stain is performed to show Gram-positive cocci in chains. Then, the organism is cultured on blood agar. The rapid pyrrolidonyl arylamidase (PYR) test is commonly used, wherein a positive reaction confers a presumptive identification of group A beta-hemolytic streptococci if the appearance and clinical context is consistent.
Gram-negative bacteria will stain a pink color due to the thin layer of peptidoglycan. If a bacteria stains purple, due to the thick layer of peptidoglycan, the bacteria is a gram-positive bacteria. [4] In clinical microbiology numerous other staining techniques for particular organisms are used (acid fast bacterial stain for mycobacteria).
As mentioned, S. agalactiae is a Gram-positive coccus with a tendency to form chains, beta-haemolytic, catalase-negative, and facultative anaerobe (anaerobic organism). GBS grows readily on blood agar plates as microbial colonies surrounded by a narrow zone of β-haemolysis.
Crystal violet stains both Gram positive and Gram negative organisms. Treatment with alcohol removes the crystal violet colour from gram negative organisms only. Safranin as counterstain is used to colour the gram negative organisms that got decolorised by alcohol. While ex vivo, many cells continue to live and metabolize until they are "fixed".
A sputum sample will also be obtained to ascertain the causative bacteria. This also plays a role in informing treatment options. [5] [6] In some cases, pulse oximetry may also be required as pneumonia is known to deplete oxygen levels in the blood. [7] Other vitals such as pulse and body temperature are also checked.
Staphylococcus saprophyticus; S. saprophyticus on Mueller–Hinton agar exhibiting resistance to novobiocin characteristic for species identification : Gram stain of a Gram positive S. saprophyticus sample suspension, lab cultured on Tryptic Soy agar (TSA)