Podzorski RP, Merline JR, Holsinger JH, Qureshi R; American Society for Microbiology. General Meeting.
Abstr Gen Meet Am Soc Microbiol. 1999 May 30-Jun 3; 99: 172 (abstract no. C-333).
Wayne State University, Detroit, MI.
Direct detection of CMV DNA in blood by PCR is frequently used to aid in the diagnosis of systemic CMV disease. Numerous procedures for processing blood specimens for PCR detection of CMV are available. In a prospective study, we compared a commercially available modified salt pre- cipitation procedure from Gentra Systems (PUREGENE Blood Kit) to an in-house developed SDS-proteinase K lysis procedure for the optimal extraction of CMV DNA from blood for subsequent PCR analysis. The PUREGENE Blood Kit utilizes whole blood while the in-house lysis procedures requires isolated peripheral blood leukocytes (PBL). The blood specimens used in this study were from bone marrow and solid organ transplant recipients being evaluated for CMV infection. 42 blood specimens were processed by both procedures and subjected to beta-globin PCR and CMV PCR. Beta-globin PCR detected amplifiable DNA in 42/42 PUREGENE Blood Kit extracts and 41/42 isolated PBL lysis extracts. 32/42 (76%) specimens were negative for CMV DNA by PCR using extracts from both procedures. 9/42 (21%) specimens were positive for CMV DNA using extracts from the PUREGENE Blood Kit. 6/42 (14%) specimens were positive for CMV DNA using extracts from the isolated PBL lysis procedure. These results suggest that the PUREGENE Blood Kit is the superior blood extraction procedure for PCR amplifiable CMV DNA when compared to an in-house developed isolated PBL lysis procedure.
Publication Types:
Keywords:
- Cytomegalovirus Infections
- DNA
- DNA Primers
- DNA, Viral
- Leukocytes
- Polymerase Chain Reaction
- Prospective Studies
- RNA-Directed DNA Polymerase
- Salts
- Sodium Chloride
- Sodium Chloride, Dietary
- blood
- methods
- transplantation
Other ID:
UI: 102195615
From Meeting Abstracts