Usefulness of ultrasensitive anti-Müllerian hormone assay to predict follicle growth in patients with primary ovarian insufficiency

Abstract Background: Patients with primary ovarian insufficiency (POI) present with follicle growth occasionally, however, it is difficult to predict the exact cycles with follicle growth. Serum anti-Müllerian hormone (AMH), a useful marker for ovarian reserve, is produced in early-stage follicles. Therefore, AMH levels should reflect the existence of small follicles which are difficult to detect using ultrasonography and may be useful for predicting follicle growth in patients with POI. Methods: Using an ultrasensitive enzyme-linked immunosorbent assay (ELISA) kit, we found very low serum AMH levels of patients with POI; we further assessed the follicle growth of each patient and their cycles to clarify the usefulness of measuring serum AMH levels for predicting the follicle growth in patients with POI. Results: A total of 91 cycles of 19 patients with POI were analyzed in this study. Five patients presented with positive serum AMH levels during the observational periods and all five experienced follicle growth. Only two of the 14 patients with negative serum AMH levels had follicle growth. Serum AMH and follicle-stimulating hormone (FSH) levels in cycles with follicle growth were significantly higher than in cycles without follicle growth. The median serum AMH level (2.77 pg/mL; 25th, 75th percentiles: 0.0, 9.64) in cycles with follicle growth were lower than the lower limit of detection of conventional AMH ELISA kits. The positive predictive value of positive serum AMH levels for follicle growth was higher than that of FSH (<10 mIU/mL).Conclusions: Measuring very low levels of serum AMH using picoAMH assays is useful for the prediction of follicle growth in patients with POI.

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Very low levels of serum anti- Müllerian hormone as a possible marker for follicle growth in patients with primary ovarian insufficiency under hormone replacement therapy

10.21203/rs.2.22083/v2 ◽

2020 ◽

Author(s):

Yukiyo Kasahara ◽

Satoko Osuka ◽

Bayasula Bayasula ◽

Natsuki Nakanishi ◽

Tomohiko Murase ◽

...

Keyword(s):

Hormone Replacement Therapy ◽

Replacement Therapy ◽

Limit Of Detection ◽

Hormone Replacement ◽

Primary Ovarian Insufficiency ◽

Enzyme Linked Immunosorbent Assay ◽

Follicle Growth ◽

Ovarian Insufficiency ◽

Positive Serum ◽

Low Levels

Abstract Background Patients with primary ovarian insufficiency (POI) occasionally present with follicle growth; however, it is difficult to predict exactly which cycles will show follicle growth. Serum anti-Müllerian hormone (AMH), a useful marker for ovarian reserve, is produced in early-stage follicles. Therefore, AMH levels should reflect the presence of small follicles, which are difficult to detect using ultrasonography, and may be useful as a marker for predicting follicle growth in patients with POI. Methods Using an ultrasensitive enzyme-linked immunosorbent assay (ELISA) kit, we found very low serum AMH levels in patients with POI. We assessed the follicle growth of each patient during each cycle to clarify the usefulness of measuring serum AMH levels for predicting follicle growth in patients with POI under hormone replacement therapy (HRT). Results A total of 91 cycles of 19 patients with POI were analyzed in this study. Five patients presented with positive serum AMH levels during the observational periods, and all five experienced follicle growth. Only two of the 14 patients with negative serum AMH levels had follicle growth. Serum AMH and follicle-stimulating hormone (FSH) levels in cycles with follicle growth were significantly higher than in cycles without follicle growth. The median serum AMH level (2.77 pg/mL; 25 th , 75 th percentiles: 0.0, 9.64) in cycles with follicle growth was lower than the lower limit of detection of conventional AMH ELISA kits. The positive predictive value of positive serum AMH levels for follicle growth was higher than that of FSH (< 10 mIU/mL). Conclusions Measurement of very low levels of serum AMH using picoAMH assays is useful for prediction of follicle growth in patients with POI under HRT conditions.

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Very low levels of serum anti-Müllerian hormone as a possible marker for follicle growth in patients with primary ovarian insufficiency under hormone replacement therapy

10.21203/rs.2.22083/v3 ◽

2020 ◽

Author(s):

Yukiyo Kasahara ◽

Satoko Osuka ◽

Bayasula Bayasula ◽

Natsuki Nakanishi ◽

Tomohiko Murase ◽

...

Keyword(s):

Hormone Replacement Therapy ◽

Replacement Therapy ◽

Limit Of Detection ◽

Hormone Replacement ◽

Primary Ovarian Insufficiency ◽

Enzyme Linked Immunosorbent Assay ◽

Follicle Growth ◽

Ovarian Insufficiency ◽

Positive Serum ◽

Low Levels

Abstract Background: Patients with primary ovarian insufficiency (POI) occasionally present with follicle growth; however, it is difficult to predict exactly which cycles will show follicle growth. Serum anti-Müllerian hormone (AMH), a useful marker for ovarian reserve, is produced in early-stage follicles. Therefore, AMH levels should reflect the presence of small follicles, which are difficult to detect using ultrasonography, and may be useful as a marker for predicting follicle growth in patients with POI. Methods: Using an ultrasensitive enzyme-linked immunosorbent assay (ELISA) kit, we found very low serum AMH levels in patients with POI. We assessed the follicle growth of each patient during each cycle to clarify the usefulness of measuring serum AMH levels for predicting follicle growth in patients with POI under hormone replacement therapy (HRT). Results: A total of 91 cycles of 19 patients with POI were analyzed in this study. Five patients presented with positive serum AMH levels during the observational periods, and all five experienced follicle growth. Only two of the 14 patients with negative serum AMH levels had follicle growth. Serum AMH and follicle-stimulating hormone (FSH) levels in cycles with follicle growth were significantly higher than in cycles without follicle growth. The median serum AMH level (2.77 pg/mL; 25th, 75th percentiles: 0.0, 9.64) in cycles with follicle growth was lower than the lower limit of detection of conventional AMH ELISA kits. The positive predictive value of positive serum AMH levels for follicle growth was higher than that of FSH (< 10 mIU/mL).Conclusions: Measurement of very low levels of serum AMH using picoAMH assays is useful for prediction of follicle growth in patients with POI under HRT conditions. Trial registration: UMIN-CTR UMIN000029464. Retrospectively registered on April 4, 2018. https://upload.umin.ac.jp/cgi-open-bin/ctr/ctr_view.cgi?recptno=R000033550

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Very Low Levels of Serum Anti-Müllerian Hormone as a Possible Marker for Follicle Growth in Patients with Primary Ovarian Insufficiency Under Hormone Replacement Therapy

Reproductive Sciences ◽

10.1007/s43032-020-00278-4 ◽

2020 ◽

Vol 28 (1) ◽

pp. 31-36

Author(s):

Yukiyo Kasahara ◽

Satoko Osuka ◽

Bayasula ◽

Natsuki Nakanishi ◽

Tomohiko Murase ◽

...

Keyword(s):

Hormone Replacement Therapy ◽

Replacement Therapy ◽

Hormone Replacement ◽

Primary Ovarian Insufficiency ◽

Follicle Growth ◽

Ovarian Insufficiency ◽

Low Levels

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Evaluation of Performance of a Commercial Monoclonal Antibody–Based Fenitrothion Immunoassay and Application to Residual Analysis in Fruit Samples

Journal of Food Protection ◽

10.4315/0362-028x-69.1.191 ◽

2006 ◽

Vol 69 (1) ◽

pp. 191-198 ◽

Cited By ~ 7

Author(s):

EIKI WATANABE ◽

KOJI BABA ◽

HEESOO EUN ◽

TOMOHITO ARAO ◽

YASUO ISHII ◽

...

Keyword(s):

Monoclonal Antibody ◽

Dynamic Range ◽

Limit Of Detection ◽

Close Correlation ◽

Gas Chromatography Mass Spectrometry ◽

Enzyme Linked Immunosorbent Assay ◽

High Recovery ◽

Elisa Kit ◽

Fruit Samples ◽

Standard Solutions

The performance of a commercially available enzyme-linked immunosorbent assay (ELISA) kit containing highly sensitive monoclonal antibodies against fenitrothion was assessed. The experimentally estimated dynamic range (0.087 to 2 ng/g) agreed with that established by the kit manufacturer (0.075 to 1 ng/g). The linearity of the standard curve produced for the kit-assembled standard solutions (slope = −0.3829) agreed with that of the curve produced for the self-made standard solutions (slope =−0.3619). The sensitivity (I50 value) and the limit of detection for the kit were 0.23 and 0.033 ng/g, respectively. Selectivity of the ELISA indicates that the monoclonal antibody can readily distinguish the target pesticide from other structurally related analogs and some metabolites (oxon forms), with the exception of ethyl O-(p-nitrophenyl) phenyl phosphonothionate (EPN), parathion-methyl, and parathion. Methanol was the best organic solvent for the kit, with optimal sensitivity observed at a final concentration in the well of 10% (vol/vol) or less. Matrix interferences were minimized by direct dilution with water (60-fold) of the methanolic extracts from apple and peach samples. To extract fenitrothion from these two agricultural products as simply and rapidly as possible, three extraction methods were used. The extraction method that involved shaking by hand for 3 min was the best among the three methods. High recovery percentages (116.6% for apple and 110.8% for peach) were obtained. Validation of the ELISA method was carried out using the gas chromatography–mass spectrometry method, resulting in high recovery and close correlation of results (r > 0.95). These findings strongly suggest that the ELISA kit may be routinely employed for on-site fenitrothion screening of fruit samples.

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Early Detection of Severe Acute Respiratory Syndrome Coronavirus 2 Antibodies as a Serologic Marker of Infection in Patients With Coronavirus Disease 2019

Clinical Infectious Diseases ◽

10.1093/cid/ciaa523 ◽

2020 ◽

Vol 71 (16) ◽

pp. 2066-2072 ◽

Cited By ~ 26

Author(s):

Zhao Rongqing ◽

Maohua Li ◽

Hao Song ◽

Jianxin Chen ◽

Wenlin Ren ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Medical Staff ◽

Early Stage ◽

Close Contact ◽

Chinese Hamster ◽

Enzyme Linked Immunosorbent Assay ◽

Elisa Kit ◽

Serologic Marker ◽

Antibody Levels ◽

Acid Test

Abstract Background Thousands of medical staff have been infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with hundreds of deaths reported. Such loss could be prevented if there were a serologic assay for SARS-CoV-2–specific antibodies for serological surveillance of its infection at the early stage of disease. Methods Using Chinese hamster ovarian (CHO) cell–expressed full-length SARS-CoV-2 S1 protein as capturing antigen, a coronavirus disease 2019 (COVID-19)/SARS-CoV-2 S1 serology enzyme-linked immunosorbent assay (ELISA) kit was developed and validated with negative samples collected prior to the outbreak or during the outbreak and positive samples from patients confirmed with COVID-19. Results The specificity of the ELISA kit was 97.5%, as examined against total 412 normal human samples. The sensitivity was 97.1% by testing against 69 samples from hospitalized and/or recovered COVID-19 patients. The overall accuracy rate reached 97.3%. The assay was able to detect SARS-CoV-2 antibody on day 1 after the onset of COVID-19 disease. The average antibody levels increased during hospitalization and 14 days after discharge. SARS-CoV-2 antibodies were detected in 28 of 276 asymptomatic medical staff and 1 of 5 nucleic acid test–negative “close contacts” of COVID-19 patients. Conclusions With the assays developed here, we can screen medical staff, incoming patients, passengers, and people who are in close contact with the confirmed patients to identify the “innocent viral spreaders,” protect the medical staff, and stop further spread of the virus.

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Serum changes in pyridinoline, type II collagen cleavage neoepitope and osteocalcin in early stage male brucellosis patients

Scientific Reports ◽

10.1038/s41598-020-72565-8 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Qiang Li ◽

Lansheng Hu ◽

Zhijun Zhao ◽

Li Ma ◽

Jiquan Li ◽

...

Keyword(s):

Early Stage ◽

Venous Blood ◽

Type Ii Collagen ◽

Serum Levels ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

Human Brucellosis ◽

Type Ii ◽

Median Serum ◽

Control Study

Abstract Musculoskeletal changes are the most common clinical manifestation of brucellosis. The main objective of this study was to provide a better understanding of this disease, while also attempting to identify potential markers that can identify the early stage musculoskeletal changes associated with human brucellosis. In this case–control study, 41 male early-stage brucellosis patients (within 6 months of diagnosis) who had not received drug therapy and 44 matched controls were examined. Venous blood samples were collected and serum pyridinoline (PYD), type II collagen cleavage neoepitope (C2C) and osteocalcin (OC) levels were quantified using an enzyme-linked immunosorbent assay (ELISA). In the brucellosis group, the median serum levels of PYD (278.53 µg/L), C2C (82.23 µg/L) and OC (8.41 µg/L) were significantly elevated relative to the control group (Z = 5.686, 3.997, 3.579; P = 0.000). Serum PYD, C2C, and OC levels were increased in early-stage male brucellosis patients, and these factors appear to have promise as potential indicator biomarkers that can reflect the osteoarticular changes that occur in the early stage of human brucellosis.

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Survey of Deoxynivalenol Contamination in Agricultural Products in the Chinese Market Using An ELISA Kit

Toxins ◽

10.3390/toxins11010006 ◽

2018 ◽

Vol 11 (1) ◽

pp. 6 ◽

Cited By ~ 6

Author(s):

Ming Li ◽

Mingna Sun ◽

Xia Hong ◽

Jinsheng Duan ◽

Daolin Du

Keyword(s):

Self Assembly ◽

Preventive Measures ◽

Limit Of Detection ◽

Agricultural Products ◽

Enzyme Linked Immunosorbent Assay ◽

Agricultural Product ◽

Chinese Market ◽

Elisa Kit ◽

Distillers Dried Grains

A total of 328 agricultural product samples highly suspected to be contaminated, from flour companies, feed companies, and livestock farms throughout China, were surveyed for deoxynivalenol (DON) contamination using a self-assembly enzyme-linked immunosorbent assay (ELISA) kit. An ELISA kit for DON was developed with a 4.9 ng mL−1 limit of detection (LOD) in working buffer and a 200 ng g−1 LOD in authentic samples. The DON contamination detection rate was 88.7%, concentrations ranged from 200.9 to 6480.6 ng g−1, and the highest DON contamination was found in distillers’ dried grains with solubles with an average of 3204.5 ng g−1. Wheat bran and wheat were found to be the most commonly contaminated samples, and the corn meal samples had the lowest average DON level. This ELISA kit is a powerful alternative method for the rapid, sensitive, specific, accurate, and high-throughput determination of DON and can meet the maximum requirement levels. This survey suggests that DON contamination in the Chinese market is serious, and the contamination risk deserves attention. Essential preventive measures should be implemented to ensure food safety and human health.

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Determination of Aflatoxin M1 in Raw Milk Using an HPLC-FL Method in Comparison with Commercial ELISA Kits—Application in Raw Milk Samples from Various Regions of Greece

Veterinary Sciences ◽

10.3390/vetsci8030046 ◽

2021 ◽

Vol 8 (3) ◽

pp. 46

Author(s):

Martha Maggira ◽

Maria Ioannidou ◽

Ioannis Sakaridis ◽

Georgios Samouris

Keyword(s):

Limit Of Detection ◽

Reference Method ◽

Raw Milk ◽

Aflatoxin M1 ◽

Enzyme Linked Immunosorbent Assay ◽

Fluorescence Detector ◽

Elisa Kit ◽

Milk Samples ◽

Of Greece

The highly toxic Aflatoxin M1 (AFM1) is most often detected in milk using an Enzyme-Linked-Immunosorbent Assay (ELISA) for screening purposes, while High-Performance Liquid Chromatography with Fluorescence Detector (HPLC-FL) is the reference method used for confirmation. The aim of the present study was the comparison between three commercially available ELISA kits and a newly developed HPLC-FL method for the determination of the AFM1 in milk samples. The developed HPLC-FL method was validated for the AFM1 and Aflatoxin M2 (AFM2), determining the accuracy, precision, linearity, decision limit, and detection capability with fairly good results. All three ELISA kits were also validated and showed equally good performance with high recovery rates. Moreover, the Limit Of Detection (LOD) and Limit Of Quantification (LOQ) values were found to be significantly lower than the Maximum Residue Limit (MRL) (50 ng kg−1). After the evaluation of all three commercial kits, the ELISA kit with the optimum performance along with the HPLC method was used for the determination of AFM1 in raw cow’s, goat’s, and sheep’s milk samples (396) obtained from producers in different regions of Greece. The evaluation of both methods showed that this ELISA kit could be considered as a faster and equally reliable alternative method to HPLC in routine analysis for the determination of AFM1 in milk.

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