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Testosterone Replacement, Low T, HCG, & Beyond
Blood Test Discussion
Why Sensitive Estradiol test?
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<blockquote data-quote="madman" data-source="post: 133521" data-attributes="member: 13851"><p>------------------------------------------------------------------------------------------------------</p><p></p><p>(direct) immunoassays:</p><p></p><p></p><p><strong><span style="color: rgb(41, 105, 176)">-they display a significant loss of specificity and accuracy at low concentrations.</span></strong></p><p></p><p><span style="color: rgb(184, 49, 47)"><strong>AND</strong></span></p><p></p><p><strong><span style="color: rgb(41, 105, 176)">-the direct immunoassays</span> <span style="color: rgb(41, 105, 176)">are prone to issues with sub-optimal</span> <span style="color: rgb(41, 105, 176)">accuracy and specificity</span> <span style="color: rgb(41, 105, 176)">due to </span><span style="color: rgb(184, 49, 47)">cross reactivity with estradiol conjugates and metabolites</span></strong></p><p></p><p></p><p></p><p></p><p><span style="font-size: 26px"><strong>Estradiol assays--The path ahead.</strong></span></p><p></p><p></p><p><span style="font-size: 18px"><strong>Abstract</strong></span></p><p>Estradiol quantitation is useful in the clinical assessment of diseases like hypogonadism, hirsutism, polycystic ovary syndrome (PCOS), amenorrhea, ovarian tumors and for monitoring response in women receiving aromatase inhibitor therapy. <strong>Physiologically relevant serum estradiol concentration in women can span across four orders of magnitude. For example, in women undergoing ovulation induction serum estradiol concentration can range between 250-2000 pg/mL whereas aromatase inhibitor therapy can decrease serum estradiol concentration to <5 pg/mL. <span style="color: rgb(184, 49, 47)">While high-through-put automated un-extracted (direct) immunoassays </span>accommodate the growing clinical need for estradiol quantitation, are amenable to implementation by most hospital clinical laboratories, <span style="color: rgb(184, 49, 47)">they display a significant loss of specificity and accuracy at low concentrations. </span></strong>Most clinical scenarios (example: estradiol monitoring in fertility treatments) place a modest demand on accuracy and precision of the assay in use <strong>but accurate quantitation of estradiol in certain clinical scenarios (pediatric and <span style="color: rgb(41, 105, 176)">male patients and for monitoring aromatase inhibitor therapy</span>)<span style="color: rgb(0, 0, 0)"> can be challenging using currently available immunoassays</span></strong> <strong><span style="color: rgb(41, 105, 176)">since the direct immunoassays</span> <span style="color: rgb(41, 105, 176)">are prone to issues with sub-optimal</span> <span style="color: rgb(41, 105, 176)">accuracy and specificity</span> <span style="color: rgb(41, 105, 176)">due to </span><span style="color: rgb(184, 49, 47)">cross reactivity with estradiol conjugates and metabolites.</span> </strong>In this review we discuss the bases for the evolution of estradiol assays from extracted (indirect) radio-immunoassays to direct immunoassays to liquid-chromatography tandem mass spectrometry (LC-MS/MS) based assays, discuss technical factors relevant for development and optimization of a LC-MS/MS assay for estradiol and present the details and performance characteristics of an ultra-sensitive LC-MS/MS estradiol assay with a limit of quantitation of 0.2 pg/mL.</p><p></p><p></p><p></p><p></p><p></p><p></p><p></p><p></p><p></p><p><strong>4. Conclusion </strong></p><p>Several studies and the recent Endocrine Society Position Statement have addressed the deficiencies in current immunoassays used for the quantitation of estradiol [2,8,9,12,13]. Direct immunoassays lack appropriate sensitivity at low estradiol levels (<40 pg/ mL). Although a GC–MS estradiol assay is considered the gold standard, it is complex to use in a routine clinical laboratory and is not amenable to high test through put. LC–MS/MS estradiol assays offer the throughput, sensitivity and precision required for estradiol quantitation over a wide range of physiological concentrations. Patients receiving AI therapy are a subset for whom high sensitivity estradiol assays are particularly useful as treatment efficacy depends on the degree of estradiol suppression. Whereas, automated direct immunoassays will remain as the first line approach for the vast majority of clinical situations where the demand for assay sensitivity is modest (example: women undergoing ovulation induction), high sensitivity LC–MS/MS assays with LOQ between 0.1 and 0.2 pg/mL are needed for optimal clinical management of certain subset of patients like those receiving AI pharmacotherapy.</p></blockquote><p></p>
[QUOTE="madman, post: 133521, member: 13851"] ------------------------------------------------------------------------------------------------------ (direct) immunoassays: [B][COLOR=rgb(41, 105, 176)]-they display a significant loss of specificity and accuracy at low concentrations.[/COLOR][/B] [COLOR=rgb(184, 49, 47)][B]AND[/B][/COLOR] [B][COLOR=rgb(41, 105, 176)]-the direct immunoassays[/COLOR] [COLOR=rgb(41, 105, 176)]are prone to issues with sub-optimal[/COLOR] [COLOR=rgb(41, 105, 176)]accuracy and specificity[/COLOR] [COLOR=rgb(41, 105, 176)]due to [/COLOR][COLOR=rgb(184, 49, 47)]cross reactivity with estradiol conjugates and metabolites[/COLOR][/B] [SIZE=26px][B]Estradiol assays--The path ahead.[/B][/SIZE] [SIZE=18px][B]Abstract[/B][/SIZE] Estradiol quantitation is useful in the clinical assessment of diseases like hypogonadism, hirsutism, polycystic ovary syndrome (PCOS), amenorrhea, ovarian tumors and for monitoring response in women receiving aromatase inhibitor therapy. [B]Physiologically relevant serum estradiol concentration in women can span across four orders of magnitude. For example, in women undergoing ovulation induction serum estradiol concentration can range between 250-2000 pg/mL whereas aromatase inhibitor therapy can decrease serum estradiol concentration to <5 pg/mL. [COLOR=rgb(184, 49, 47)]While high-through-put automated un-extracted (direct) immunoassays [/COLOR]accommodate the growing clinical need for estradiol quantitation, are amenable to implementation by most hospital clinical laboratories, [COLOR=rgb(184, 49, 47)]they display a significant loss of specificity and accuracy at low concentrations. [/COLOR][/B]Most clinical scenarios (example: estradiol monitoring in fertility treatments) place a modest demand on accuracy and precision of the assay in use [B]but accurate quantitation of estradiol in certain clinical scenarios (pediatric and [COLOR=rgb(41, 105, 176)]male patients and for monitoring aromatase inhibitor therapy[/COLOR])[COLOR=rgb(0, 0, 0)] can be challenging using currently available immunoassays[/COLOR][/B] [B][COLOR=rgb(41, 105, 176)]since the direct immunoassays[/COLOR] [COLOR=rgb(41, 105, 176)]are prone to issues with sub-optimal[/COLOR] [COLOR=rgb(41, 105, 176)]accuracy and specificity[/COLOR] [COLOR=rgb(41, 105, 176)]due to [/COLOR][COLOR=rgb(184, 49, 47)]cross reactivity with estradiol conjugates and metabolites.[/COLOR] [/B]In this review we discuss the bases for the evolution of estradiol assays from extracted (indirect) radio-immunoassays to direct immunoassays to liquid-chromatography tandem mass spectrometry (LC-MS/MS) based assays, discuss technical factors relevant for development and optimization of a LC-MS/MS assay for estradiol and present the details and performance characteristics of an ultra-sensitive LC-MS/MS estradiol assay with a limit of quantitation of 0.2 pg/mL. [B]4. Conclusion [/B] Several studies and the recent Endocrine Society Position Statement have addressed the deficiencies in current immunoassays used for the quantitation of estradiol [2,8,9,12,13]. Direct immunoassays lack appropriate sensitivity at low estradiol levels (<40 pg/ mL). Although a GC–MS estradiol assay is considered the gold standard, it is complex to use in a routine clinical laboratory and is not amenable to high test through put. LC–MS/MS estradiol assays offer the throughput, sensitivity and precision required for estradiol quantitation over a wide range of physiological concentrations. Patients receiving AI therapy are a subset for whom high sensitivity estradiol assays are particularly useful as treatment efficacy depends on the degree of estradiol suppression. Whereas, automated direct immunoassays will remain as the first line approach for the vast majority of clinical situations where the demand for assay sensitivity is modest (example: women undergoing ovulation induction), high sensitivity LC–MS/MS assays with LOQ between 0.1 and 0.2 pg/mL are needed for optimal clinical management of certain subset of patients like those receiving AI pharmacotherapy. [/QUOTE]
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Testosterone Replacement, Low T, HCG, & Beyond
Blood Test Discussion
Why Sensitive Estradiol test?
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