DNA Sequencing Reveals Another Novel Lyme Disease Spirochete in Blood of Boy Diagnosed with Psychiatric Disorders, Reports Connecticut Pathologist Sin Hang Lee, M.D.

Standard Lyme disease test kits cannot diagnose as yet unrecognized
borrelial strains of Lyme, says Dr. Lee

HARTFORD, Conn.–(BUSINESS WIRE)–During the winter month of February 2014, Sin Hang Lee, M.D., of Milford
Molecular Diagnostics laboratory in Milford, Conn., detected an unusual
strain of Borrelia burgdorferi with two homeologous 16S
rRNA genes by DNA sequencing in the blood of a boy discharged from a
psychiatric hospital. This is the second novel strain of borrelia
detected in a patient’s blood samples by Dr. Lee, as reported in an
April 21, 2016 article in International Medical Case
Reports Journal (https://www.dovepress.com/article_26575.t53729944).

As reported, a teenager living in Massachusetts was initially diagnosed
with Lyme disease and treated with a full 28-day course of antibiotics.
However, he subsequently developed a variety of peculiar symptoms, the
severity of which prevented him from attending school for a year. A Lyme
disease consultant, called in to review the case, found the serology
test results non-diagnostic of Lyme disease. Based on the consultant’s
opinion, the patient was then hospitalized for pure psychiatric
disorders at a psychiatric hospital for seven weeks. After discharge
from the psychiatric hospital, a venous blood sample taken from the boy
by his primary care physician was tested positive for Borrelia
burgdorferi by 16S rRNA gene sequencing. The patient was then
referred to a major general medical center in Boston for treatment.
Further analysis of the sequencing data revealed that the Borrelia
burgdorferi detected in this case has two partially homologous
(homeologous) 16S rRNA genes, a hitherto unrecognized gene organization
in Lyme disease spirochetes.

In a separate instance, occurring approximately two years ago, Dr. Lee
reported uncovering another novel strain of borrelia through DNA
sequencing in an archived serum sample from a patient from another
state. The patient had been treated for neurologic Lyme disease, also by
16S rRNA gene sequencing http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3975398/.

Each bacterial species has its unique 16S rRNA gene needed for
initiating protein synthesis and for life maintenance. Researchers using
pure bacterial cultures as the study materials in the past showed that
all spirochete isolates from Lyme disease patients in North America
belong to the species of Borrelia burgdorferi sensu stricto and
all have only one copy of 16S rRNA gene. The commercial serology test
kits for Lyme disease are designed for the diagnosis of infections
caused by this one species of Lyme disease spirochetes. However, other
uncultivatable borrelial strains with different genetic makeups can also
cause Lyme disease but with atypical serology patterns which are
considered non-diagnostic by the criteria promulgated by these
diagnostic test kits, as Dr. Lee explained in this case report.

In the current report, Dr. Lee said, “Using a pair of genus-specific PCR
primers to perform same-nested PCR for amplification of the borrelial
16S rRNA gene, followed by direct Sanger sequencing of the PCR amplicon,
can detect and validate a wide range of Borrelia species, including
novel Borrelia strains causing Lyme and related borrelioses. Direct DNA
sequencing should be implemented in hospital laboratories in Lyme
disease-endemic areas for early reliable diagnosis of this infectious
disease and for further studies of the diversity of the causative agents
of Lyme borreliosis.”

“People living in or visiting Lyme disease endemic areas should be aware
of possible infections by not yet recognized borrelial strains which
cannot be diagnosed by the standard test kits,” said Dr. Lee.

Dr. Lee, who has been practicing pathology in New Haven County
Connecticut since 1971, is the director of Milford Molecular Diagnostics
laboratory in Milford, Connecticut (http://www.dnalymetest.com/).
He has been using DNA sequencing for molecular diagnosis of Lyme and
related borrelioses since 2009 when he was a pathologist at Milford
Hospital.

Contacts

For Milford Molecular Diagnostics Laboratory
Kevin Moore,
203-788-8497