AJCP / Meeting Abstracts

Detection of Putative Novel Mutation in Zp1 Gene J.S. Hizon1, J.C. Macalalad1, R.G. Sison1, X.A. Nieto2;
1
Causing Empty Follicle Syndrome Department of Medical Technology, University of Santo
D. Upadhyay1, S. Anandt1, B. Peramo2, H. Al Safar3; Tomas Manila, Manila, Metro Manila, Philippines,
2
Genetics Department, Al Ain Fertility Center, Al Ain, College of Science, Department of Mathematics and
Abu Dhabi, United Arab Emirates, 2Medical Director, Statistics, University of Santo Tomas Manila, Manila,
Al Ain Fertility Center, Al Ain, Abu Dhabi, United Arab Metro Manila, Philippines
Emirates, 3Center for Biotechnology, Khalifa University,
Abu Dhabi, Abu Dhabi, United Arab Emirates Introduction/Objective:  Continuous progression of
the COVID-19 outbreak prompted an expensive, lim-
Introduction/Objective: Mutations in the zona pellucida ited, and slow-moving laboratory-based molecular
glycoprotein genes have been reported to be associated testing capacity of the gold standard, Real-time Reverse

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with empty follicle syndrome (EFS) and abnormal zona Transcription- Polymerase Chain Reaction (rRT-PCR),
pellucida (ZP). At our clinic, in a cohort of infertile pa- in detecting SARS-CoV-2. The vital step in mitigation is
tients, we found three cases with empty follicle syndrome utilizing an accurate diagnostic detection assay to deploy
as a root cause of infertility. Further, we designed a study various preventive measures. Therefore, there is a need
to identify the disease-causing gene mutations in these to evaluate an accessible, fast, and inexpensive point-
patients. At present, there are no citations or functional of-care immunoassay, such as Antigen-detecting Rapid
evidence in ClinVar for NM_207341.3:c.1168del variant Diagnostic Tests (Ag- RDTs). Thus, the study aimed to
indicating the involvement of this variant in empty fol- assess the diagnostic accuracy of Ag-RDT in detecting
licle syndrome. But it has been interpreted to be Likely SARS-CoV-2.
pathogenic causing female infertility due to zona pellu- Methods/Case Report: A non-experimental, quantita-
cida defect. tive, and cross-sectional cohort study utilized 272 SARS-
Methods/Case Report: In this prospective study con- CoV-2 Ag-RDT with rRT-PCR confirmation collected
ducted at Al Ain Fertility Center (AAFC), U.A.E, we from April 2021 to February 2022 in a private primary
performed genetic analysis on three patients (first-degree hospital laboratory database in Bulacan, Philippines.
sisters) having a mean age of 31.7 (2.52) years. Patient Diagnostic accuracy parameters of Ag-RDT included
indications were primary female infertility and IVF in the study are sensitivity, specificity, positive predictive
failures due to no eggs present in the follicles when egg value (PPV), and negative predictive value (NPV).
retrieval was attempted. We characterized these three pa- Results (if a Case Study enter NA):  The overall sensitivity
tients as suffering from empty follicle syndrome. Along and specificity of Ag-RDT were 69.6% and 98.8%, re-
with the patient's sample, we also included their common spectively. Additionally, it exhibited a PPV of 84.2% and
parent's sample for Whole Exome Sequencing and Sanger an NPV of 97.2%. With a hypothetical 1% prevalence,
sequencing, to rule out the inheritance pattern in the PPV and NPV were 36.8% and 99.7%. Meanwhile, PPV
family. The effects of this mutation were further charac- and NPV were 86.5% and 96.7% at 10% prevalence. High
terized through mRNA and protein studies. specificity denotes that positive Ag-RDT test results can
Results (if a Case Study enter NA): N/A. be confidently considered in true SARS-CoV-2 infection,
Conclusion:  To our knowledge, this is the first study in the while low sensitivity entails that a negative result must be
U.A.E. showing the familial novel variant in the ZP1 gene subjected to confirmatory tests. Furthermore, when prev-
associated with empty follicle syndrome. We identified a alence increases, PPV increases while NPV decreases.
novel mutation in these patients (NM_207341.3:c.1168del) Conclusion: The study primarily encapsulated that the
in the ZP1 gene in the homozygous state and the same Ag-RDT cannot replace rRT-PCR in detecting SARS-
variant in the heterozygous state in the parents. Our find- CoV-2 infections. Despite its shortcomings, Ag-RDT is
ings add to the existing mutational spectrum of the ZP1 still valuable in areas with moderate-to-high disease prev-
gene associated with empty follicle syndrome and ab- alence and with limited access to confirmatory test due to
normal oocytes and provide new support for the genetic its high predictive values and advantages. A larger number
diagnosis of female infertility. of positive results of rRT-PCR, evaluation of factors cor-
relating to low sensitivity, and a prospective study are re-
commended for future studies.

The Assessment of the Diagnostic Accuracy of Antigen-

Detecting Rapid Diagnostic Test using Nasopharyngeal
Swab Specimen in Detecting SARS-CoV-2 among Analysis of microRNAs by the Stem-Loop Real-Time
Patients in a Private Hospital in Bulacan, Philippines Quantitative Reverse Transcription Polymerase Chain
Reaction Using A Double Quencher Probe
R.M. Magpoc1, M. Murzo1, A.P. Jayag1, S.P. Lee1,
A.M. Manguiat1, E.M. Oloteo1, A. Mendoza1, A. Kikuchi1, A. Naruse1, K. Takagi2; 1Research
Institute for Medical Science, Daiyukai Health System,

S142 Am J Clin Pathol 2022;158:S30-S161 © American Society for Clinical Pathology

DOI: 10.1093/ajcp/aqac126
 AJCP  / Meeting Abstracts

Ichinomiya, Aichi, Japan, 2Department of Urology, patients are asymptomatic which can be a challenge in
Daiyukai Daiichi Hospital, Ichinomiya, Aichi, Japan controlling the spread of disease. Antibody detection
by chemiluminescence immunoassay (CLIA) is a good
Introduction/Objective:  We created a universal probe that complementary test to Real-Time Reverse Transcription
can be used with mature micro RNAs (miRNAs) using a – Polymerase Chain Reaction (rRT-PCR) as an indirect
double quencher incorporating the ZEN quencher into the marker for disease.
hydrolysis probe required for miRNA analysis by the real- Methods/Case Report:  This retrospective study assessed
time quantitative polymerase chain reaction (RT-qPCR) the clinico-demographic characteristics of 184 SARS-
using a stem-loop primer. The main purpose of this ap- CoV-2 IgM and IgG antibody-reactive patients at Gov.
proach was to improve sensitivity and convenience. Celestino Gallares Memorial Hospital from July 1, 2020 –
Methods/Case Report:  We conducted studies on miR-21, May 31, 2021.

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miR-30a and miR-200c. To assess the relationship be- Results (if a Case Study enter NA):  A total of 184 cases
tween the sequence interval and reactivity between the were included in the study out of 4,138 patients tested
miRNA-specific sequence on the stem-loop primer and for anti-SARS-CoV-2 IgM and IgG antibodies by CLIA.
detection probe, various sequence patterns were examined Within this population, 97 were outpatients and 87 were
using miRNA from K562 cells. The sensitivity and reac- inpatients. Majority of patients needing hospitalization
tivity of the designed primers and probes were examined were aged ≥60-year-old males (33.33%) and that the prob-
using each synthetic miRNA and transurethral resection ability of being admitted decreases with younger age.
of bladder tumor (TURBT) samples. The synthetic RNA Most of the outpatients who were reactive to antibodies
was serially diluted in 10% increments. Twenty-seven against SARS-CoV-2 were frontline workers compared to
TURBT cases were examined. The prepared samples non-frontline workers and non-workers (27.84% versus
were extracted with the FastGene RNA Basic Kit and 1.03% and 19.59%). Most common comorbidities were
FastGene miRNA Enhancer. After extraction, cDNA hypertension (24.46%), cardiovascular disease (15.76%),
was prepared using FastGene Scriptase II. We performed diabetes mellitus (11.41%), and chronic kidney disease
quantitative analysis by the RT-qPCR using a ZEN (11.41%). All five mortalities were diagnosed hypertensive
double quencher probe. The samples were analyzed with who later developed critical disease.
the Taq Man miRNA assay. Conclusion:  The continued use of antibody detection by
Results (if a Case Study enter NA): The probe and CLIA as a complementary test with rRT-PCR is still re-
miRNA-specific sequences of the stem-loop primer commended not only for better detection of COVID-19
showed better reactivity when the design contained less especially in asymptomatic patients but also to observe
space within the sequences. Serial dilution of the samples changes in antibody titer especially in the advent of
using synthetic miRNA showed that linearity depended COVID-19 vaccines.
on the addition amount, and occurred at a concentration
of log 7 or more. In the TURBT samples, the Δ of the
mean Cp value, and the correlation coefficients of miR-
21, miR-30a, and miR-200c, were 3.13 and 0.966; 1.10 Rapid Detection of Uropathogens and Antibiotic-resistant
and 0.991; and 0.39 and 0.997, respectively, compared Genes Using Open Array Multiplex PCR Technology
with the TaqMan miRNA assay. P. Neopane1, J. Nypaver1, S.S. Beqaj1, R. Shrestha2;
Conclusion:  This demonstrates that our method has the 1
Molecular, Patients Choice Laboratories, Zionsville,
same or higher sensitivity than the TaqMan miRNA Indiana, United States, 2Patients Choice Laboratories,
assay. Overall, these findings suggest that the analysis Indianapolis, Indiana, United States
of miRNA with a stem-loop primer using the double
quencher probe is useful. Introduction/Objective:  Introduction: Standard diag-
nostic practices for Urinary Tract Infection (UTI) take
2–3  days for pathogen identification and antibiotic sus-
ceptibility testing leading to delayed diagnosis and an in-
Clinico-Demographic Characteristics of Patients with crease in healthcare expenditures. The rapid diagnostics
SARS-CoV-2 Antibodies: A Single Tertiary Center tool is highly desirable to identify the pathogen and its
Retrospective Study in Bohol, Philippines antibiotic-resistant genes directly from the samples. Our
A.L. Salillas, A.B. Clapis, E.G. Almocera, A.T. San Juan; objective is to validate and evaluate the performance of
Pathology, Gov. Celestino Gallares Memorial Hospital, Open array multiplex PCR for pathogen identification
Tagbilaran, Bohol, Philippines and antibiotic resistance genes.
Methods/Case Report: Method: Open array multi-
Introduction/Objective: Coronavirus Disease 2019 plex PCR method for 20 UTI pathogens was validated
(COVID-19) caused by SARS-CoV-2 continues to be using ATCC organisms and inter-laboratory corre-
a global threat until the present. Most of the infected lation using 50 urine samples. Briefly, samples were

© American Society for Clinical Pathology Am J Clin Pathol 2022;158:S30-S161 S143

DOI: 10.1093/ajcp/aqac126