Selectively & sensitively detect viable bacteria - PMA™ dyes & kits

These novel dyes from Biotium enable selective and sensitive detection of viable bacteria in the presence of dead bacteria. Traditional bacterial culture is time-consuming and has low sensitivity, whereas detection based on PCR is rapid and highly sensitive but cannot distinguish live from dead cells. Treating bacteria with propidium monoazide (PMA™) dye prior to PCR analysis allows selective detection of only viable bacteria in a highly sensitive and reliable manner. These dyes have been widely referenced on the use of the dye in pathogenic bacterial detection related to food and water safety, medical diagnosis and biodefence.

PMA™ dye
The DNA-modifying PMA™ dye is weakly fluorescent, becoming more fluorescent after binding to dsDNA with high affinity. Upon photolysis, the dye’s photoreactive azido group is converted to a highly reactive nitrene radical, which readily reacts with any hydrocarbon moiety at the binding site to form a stable covalent nitrogen-carbon bond, thus resulting in permanent DNA modification. The dye is almost completely cell membrane-impermeable so can be selectively used to modify only exposed DNA from dead cells while leaving DNA from viable cells intact. This feature makes the dye highly useful in the selective detection of viable pathogenic cells by quantitative real-time PCR in the presence dead cells whose DNA has been PMA-modified and thus cannot be amplified.
 


Cell membrane-impermeable PMA dye selectively and covalently modifies DNA from dead bacteria with compromised membrane while leaving DNA from viable cells intact. Because PMA-modified DNA cannot be amplified, subsequent lysis of viable cells and qPCR permit selective quantitation of viable bacteria.

Benefits of using the PMA™ dye
• Selectively detect live from dead cells
• Highly sensitive
Representative real-time PMA-PCR amplification curves from untreated and heat-killed E. coli, using primers against 16S rRNA.
 


The delta Ct of live and killed E. coli with and without PMA treatment. The Ct value of sample without PMA was subtracted from the corresponding sample with PMA cross-linking (Ct with PMA – Ct without PMA)

PMA™ real-time PCR bacterial viability kits
PMA™-PCR kits are designed for selective detection of viable bacteria from a specific strain using PMA™ dye and real-time PCR. The kits contain PMA™ dye, Fast EvaGreen® qPCR Master Mix, and validated PCR primers for detection of selected strains of bacteria that are of widespread interest to food safety, public health and/or antibacterial research.

Kits include:
• PMA dye OR PMAxx dye
• Fast EvaGreen qPCR Master Mix
• ROX reference dye
• PMA Enhancer (gram-negative strains only)
• Validated PCR primers for specific bacterial strain

PMA™ real-time PCR bacterial viability kits are available for:
• Salmonella enterica
• Mycobacterium tuberculosis
• Staphylococcus aureus
• Staphylococcus aureus (methicillin-resistant)
• Listeria monocytogenes
• E. coli
• E. coli O157:H7
• Legionella pneumophila

Material available for download
PMA™ Dye Protocol
PMA™ Dye MSDS
PMA Dye (20 mM in H2O) MSDS
PMAxx Protocol
PMAxx MSDS
PMA Real-Time PCR Bacterial Viability Kit – Staphylococcus aureus (mecA) Protocol
PMA Real-Time PCR Bacterial Viability Kit – Staphylococcus aureus (mecA) MSDS
PMA Real-Time PCR Bacterial Viability Kit – Staphylococcus aureus (nuc) Protocol
PMA Real-Time PCR Bacterial Viability Kit – Staphylococcus aureus (nuc) MSDS
PMA Real-Time PCR Bacterial Viability Kit – Salmonella enterica (invA) Protocol
PMA Real-Time PCR Bacterial Viability Kit – Salmonella enterica (invA) MSDS
PMA Real-Time PCR Bacterial Viability Kit – Mycobacterium tuberculosis (groEL2) Protocol
PMA Real-Time PCR Bacterial Viability Kit – Mycobacterium tuberculosis (groEL2) MSDS
PMA Real-Time PCR Bacterial Viability Kit – Listeria monocytogenes (hly) Protocol
PMA Real-Time PCR Bacterial Viability Kit – Mycobacterium tuberculosis (groEL2) MSDS
PMA Real-Time PCR Bacterial Viability Kit – Legionella pneumophila (mip) Protocol
PMA Real-Time PCR Bacterial Viability Kit – Legionella pneumophila (mip) MSDS
PMA Real-Time PCR Bacterial Viability Kit – E. coli O157:H7 (Z3276) Protocol
PMA Real-Time PCR Bacterial Viability Kit – E. coli O157:H7 (Z3276) MSDS
PMA Real-Time PCR Bacterial Viability Kit – E. coli (uidA) Protocol
PMA Real-Time PCR Bacterial Viability Kit – E. coli (uidA) MSDS
Viability PCR With PMA & PMAxx™ Dyes Flyer

PMA Reference List

Products

Note: product availability depends on country - see product detail page.

Details Cat number & supplier Size Price
PMA BT40013 · Biotium
BT40013
Biotium
1 mg £150.00
1 mg
view
PMA, 20mM in H2O BT40019 · Biotium
BT40019
Biotium
100 µl £144.00
100 µl
view
PMAxx™, 20mM in H2O BT40069 · Biotium
BT40069
Biotium
100 µl £144.00
100 µl
view
PMA Real-Time PCR Bacterial Viability Kit - Staphylococcus aureus (mecA) BT31036 · Biotium
BT31036
Biotium
1 kit £357.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - Staphylococcus aureus (nuc) BT31035 · Biotium
BT31035
Biotium
1 kit £357.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - Salmonella enterica (invA) BT31033 · Biotium
BT31033
Biotium
1 kit £386.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - Salmonella enterica (invA) PMAxx™ BT31033-X · Biotium
BT31033-X
Biotium
1 kit £386.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - Mycobacterium tuberculosis (groEL2) BT31034 · Biotium
BT31034
Biotium
1 kit £357.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - Listeria monocytogenes (hly) BT31051 · Biotium
BT31051
Biotium
1 kit £357.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - Listeria monocytogenes (hly) PMAxx™ BT31051-X · Biotium
BT31051-X
Biotium
1 kit £386.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - Legionella pneumophila (mip) BT31053 · Biotium
BT31053
Biotium
1 kit £386.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - E. coli O157:H7 (Z3276) BT31037 · Biotium
BT31037
Biotium
1 kit £386.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - E. coli O157:H7 (Z3276) PMAxx™ BT31037-X · Biotium
BT31037-X
Biotium
1 kit £386.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - E. coli (uidA) BT31050 · Biotium
BT31050
Biotium
1 kit £386.00
1 kit
view
PMA Real-Time PCR Bacterial Viability Kit - E. coli (uidA) PMAxx™ BT31050-X · Biotium
BT31050-X
Biotium
1 kit £386.00
1 kit
view