Molecular analysis of pathogenic bacteria and fungi in liquid samples from clinical materials and animal systems (e.g., blood and other body liquids) can be severely disturbed by a high background of host and dead cell DNA. Besides PCR inhibitors, unspecific binding of bacterial and fungal sequence-specific primers to host sequences can negatively interfere with pathogen analysis. With MolYsis™ Complete5 Molzym has developed a tool for the enrichment of bacterial and fungal cells and isolation of pure microbial DNA from blood and other liquids. MolYsis™ Complete5 is the complete solution for the removal of PCR inhibitors and host and dead cell DNA from samples, allowing the reliable and sensitive detection of microbial pathogens through PCR or Real-Time PCR.
MolYsis™ Complete5 allows for the microbial DNA isolation from human and animal liquid samples:
i) Samples of 1ml from pediatric patients.
ii) Samples of 5ml from adult patients.
Whole blood (with anti-coagulants), synovial fluid, pleural fluid, cerebrospinal fluid, ascites fluid, pus, broncho-alveolar lavage, nasal douche fluid, urine.
Whole blood (with anti-coagulants) from mouse, rat, and monkey, hamster ovary cell culture (≤5*108 cells per sample), monkey renal cell culture.
Only three steps are needed to obtain microbial DNA preparations that are depleted of host DNA (Fig. 1):
I) The addition of a chaotropic buffer to a liquid sample lyses the host cells, whereas microbial cells are unaffected. The DNA released from host cells as well as dead cells is degraded by Molzym’s proprietary, chaotrope-resistant MolDNase B.
II) Microbial cells are sedimented, treated with BugLysis reagents to degrade cell walls of Gram-negative bacteria, Gram-positive bacteria and fungi and then digested by Proteinase K treatment.
III) The microbial DNA is extracted and then isolated by a quick bind-wash-elute procedure, using Molzym’s CCT technology with quantitative DNA binding to the filter matrix and high recovery of microbial DNA from the column.
Fig. 1: The principle of testing for bacterial and fungal DNA in samples by MolYsis™ Complete5.
APPLICATIONS MOLYSIS COMPLETE 10:
– Sepsis diagnosis of adult patients (10 ml protocol)
– Sepsis diagnosis of adult patients (5 ml protocol)
– Sepsis diagnosis of child patients (1 ml protocol)
– Infection diagnosis of other body fluids, including cerebrospinal fluids and joint aspirates
- MolDNase (DNA degradation)
- BugLysis (degradation of Gram-positive and Gram-negative cell walls)
- Mini spin columns
- 10.0 ml blood (extra protocol for 1.0 ml and 5.0 ml included)
APPLICATION NOTE: MOLYSIS – MICROBIAL DNA ISOLATION FROM DIVERSE CLINICAL SPECIMENS FOR SENSITIVE MOLECULAR DIAGNOSIS.
Ordering Information MolYsis (click on the product for manual)
|Molysis Basic||D-300-050||50 reactions|
|Molysis Basic||D-300-100||100 reactions|
|Molysis Basic 5||D-301-050||50 reactions|
|Molysis Basic 5||D-301-100||100 reactions|
|Molysis Basic 10||D-305-050||50 reactions|
|Molysis Basic 10||D-305-100||100 reactions|
|Molysis Complete 5||D-321-050||50 reactions|
|Molysis Complete 5||D-321-100||100 reactions|
|Molysis Complete 10||D-325-050||50 reactions|
|Molysis Complete 10||D-325-100||100 reactions|
|Molysis Plus||D-310-050||50 reactions|
|Molysis Plus||D-310-100||100 reactions|