The Role of Culture and Nucleic Acid Amplification in Diagnosis of Lyme Borreliosis

Wormser, Gary P.; Wang, Guiqing

doi:10.1002/9780470933961.ch9

Lyme Borreliosis in Europe and North America 2011

DOI: 10.1002/9780470933961.ch9

|View full text |Cite

The Role of Culture and Nucleic Acid Amplification in Diagnosis of Lyme Borreliosis

Gary P. Wormser

Guiqing Wang

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...

Citation Types

Supporting

Mentioning

Contrasting

Year Published

2013

Publication Types

Select...

Article1

Relationship

Self Cite0

Independent1

Authors

Journals

Cited by 1 publication

(1 citation statement)

References 159 publications

(200 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A bility to culture Borrelia burgdorferi has been essential for clinical, epidemiological, and pathogenicity studies of B. burgdorferi (1). Various culture medium formulations for Borrelia growth have been widely used to isolate B. burgdorferi from ticks, hosts, and clinical samples (2)(3)(4).…”

mentioning

confidence: 99%

Detection of Heterogeneity of Borrelia burgdorferi in Ixodes Ticks by Culture-Dependent and Culture-Independent Methods

Wesker

White

et al. 2013

J Clin Microbiol

View full text Add to dashboard Cite

bAbilities to detect heterogeneity of ospC genotypes of the Lyme disease spirochete Borrelia burgdorferi in the tick vector by in vitro culture (IVC) and direct PCR (dPCR) were compared. IVC failed to detect one-third of the ospC genotypes detected by dPCR. Among IVC results, common ospC genotypes were overrepresented while occurrence of rare genotypes was underestimated.A bility to culture Borrelia burgdorferi has been essential for clinical, epidemiological, and pathogenicity studies of B. burgdorferi (1). Various culture medium formulations for Borrelia growth have been widely used to isolate B. burgdorferi from ticks, hosts, and clinical samples (2-4). More recently, culture-independent means of Borrelia detection, usually involving PCR, have become the gold standard (5). One widely used genetic marker for detection of B. burgdorferi is the gene encoding outer surface protein C (ospC). The ospC gene encodes an important major surface protein that has proven useful for discriminating several genotypes (6-9).A comparison of B. burgdorferi strains cultured from ticks with those cultured from clinical samples has led to conclusions that a limited set of ospC genotypes (A, B, I, and K) are invasive and cause systemic disease while the others are limited to ticks or persist as localized infections (8, 10). While several investigators have reported a cultivation effect of B. burgdorferi, all results are based on clinical samples and/or conserved genetic markers. In this study, we compared performances of direct PCR (dPCR) and in vitro cultivation (IVC) to detect 17 distinct Borrelia burgdorferi ospC genotypes from tick specimens.A total of 926 adult deer ticks were surface sterilized in providone-iodine and then rinsed in 70% EtOH. Ticks in one treatment group (n ϭ 420) were bisected by a dorsoventral medial cut, and one half of the ticks were placed directly into BSK-H medium (Sigma-Aldrich, St. Louis, MO) while the other half were used for DNA extraction. Ticks in a second treatment group (n ϭ 506) were dissected, and gut contents were placed in 1.5 ml BSK-H medium; the remainder of the tick body was used in DNA extraction as described above. All cultures were incubated at 34°C and checked biweekly for 4 weeks by dark-field microscopy.After 4 weeks, 25 l of each culture was transferred to a 96-well plate containing 75 l H 2 O, heated to 95°C for 10 min, and centrifuged at 6,000 ϫ g for 10 min, and the resulting supernatant was used as a template for PCR. DNA extraction was performed using the Epicentre Master Complete DNA and RNA purification kits (Epicentre Technologies, Madison, WI). PCR conditions for ospC and the reverse line blotting technique were as described previously (7,11). In addition to the 15 published probes, two new ospC probes were used in this study. We named these type-V (5=-GAG CCG CTT GAG CAG TTA AAC CAT TTG CAC C-3=) and type-W (5=-TCG TTT CGA TTT GCT TCT ACA CCC-3=).dPCR of ticks detected a higher overall infection rate (61.9%) than IVC (34.3%) ( 2 ϭ 140.64; degrees of freedom [df] ϭ 1; P Ͻ...

show abstract

mentioning

confidence: 99%

Detection of Heterogeneity of Borrelia burgdorferi in Ixodes Ticks by Culture-Dependent and Culture-Independent Methods

Wesker

White

et al. 2013

J Clin Microbiol

View full text Add to dashboard Cite

show abstract

scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.

Contact Info

hi@scite.ai

10624 S. Eastern Ave., Ste. A-614

Henderson, NV 89052, USA

Blog Terms and Conditions API Terms Privacy Policy Contact Cookie Preferences Do Not Sell or Share My Personal Information

Made with 💙 for researchers

Part of the Research Solutions Family.

The Role of Culture and Nucleic Acid Amplification in Diagnosis of Lyme Borreliosis

Cited by 1 publication

References 159 publications

Detection of Heterogeneity of Borrelia burgdorferi in Ixodes Ticks by Culture-Dependent and Culture-Independent Methods

Detection of Heterogeneity of Borrelia burgdorferi in Ixodes Ticks by Culture-Dependent and Culture-Independent Methods

Contact Info

Product

Resources

About