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Diagnosis of Balamuthia mandrillaris Encephalitis by Thymine-Adenine Cloning Using Universal Eukaryotic Primers

Authors
 Ju Yeong Kim  ;  Myung-Hee Yi  ;  Myungjun Kim  ;  Joon-Sup Yeom  ;  Hyun Dong Yoo  ;  Seong Min Kim  ;  Tai-Soon Yong 
Citation
 ANNALS OF LABORATORY MEDICINE, Vol.42(2) : 196-202, 2022-03 
Journal Title
ANNALS OF LABORATORY MEDICINE
ISSN
 2234-3806 
Issue Date
2022-03
MeSH
Adenine ; Balamuthia mandrillaris* / genetics ; Cloning, Molecular ; Encephalitis* / diagnosis ; Eukaryota ; Humans ; Male ; Middle Aged ; Thymine
Keywords
18S rRNA ; Amoeba ; Balamuthia mandrillaris ; Encephalitis ; TA cloning
Abstract
Background: Identifying the causal pathogen of encephalitis remains a clinical challenge. A 50-year-old man without a history of neurological disease was referred to our department for the evaluation of an intracranial lesion observed on brain magnetic resonance imaging (MRI) scans, and the pathology results suggested protozoal infection. We identified the species responsible for encephalitis using thymine-adenine (TA) cloning, suitable for routine clinical practice.

Methods: We extracted DNA from a paraffin-embedded brain biopsy sample and performed TA cloning using two universal eukaryotic primers targeting the V4-5 and V9 regions of the 18S rRNA gene. The recombinant plasmids were extracted, and the inserted amplicons were identified by Sanger sequencing and a homology search of sequences in the National Center for Biotechnology Information Basic Local Alignment Search Tool.

Results: The infection was confirmed to be caused by the free-living amoeba Balamuthia mandrillaris. Two of 41 colonies recombinant with 18S V4-5 primers and 35 of 63 colonies recombinant with the 18S V9 primer contained B. mandrillaris genes; all other colonies contained human genes. Pathogen-specific PCR ruled out Entamoeba histolytica, Naegleria fowleri, Acanthamoeba spp., and Toxoplasma gondii infections.

Conclusions: This is the first report of B. mandrillaris-induced encephalitis in Korea based on molecular identification. TA cloning with the 18S rRNA gene is a feasible and affordable diagnostic tool for the detection of infectious agents of unknown etiology.
Files in This Item:
T202202892.pdf Download
DOI
10.3343/alm.2022.42.2.196
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Tropica Medicine (열대의학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Ju Yeong(김주영) ORCID logo https://orcid.org/0000-0003-2456-6298
Yeom, Joon Sup(염준섭) ORCID logo https://orcid.org/0000-0001-8940-7170
Yong, Tai Soon(용태순) ORCID logo https://orcid.org/0000-0002-3445-0769
Yi, Myung Hee(이명희) ORCID logo https://orcid.org/0000-0001-9537-5726
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/189529
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