Full specifications for every recommended test — what it measures, which cascade node it confirms, where to order it, and when.
In regressive presentations, tests marked HIGH in the regression column should be prioritized immediately regardless of standard sequencing. These markers reflect active system instability rather than background dysfunction.
This document is the lookup companion to Document 07 — Where to Start. If you have not yet identified which channels are most relevant to your child's presentation, begin there. This reference is for when you know which direction you are headed and need full test specifications: what a test measures, which cascade node it confirms, where to order it, and how it is prioritized within the channel investigation sequence.
First-line tests are ordered routinely for all individuals investigating that channel — they give the broadest directional signal. Second-line tests are ordered when first-line results confirm the channel is active and you need to quantify severity or narrow the mechanism. Third-line tests are for specific circumstances, unexplained findings, or when precision mapping of a confirmed channel is required for intervention design.
All tests in this reference are used to identify which upstream biological channels are active in a given individual — not to diagnose autism. Abnormal findings guide the reordering of the intervention protocol. They do not confirm or deny the diagnosis. All testing decisions require clinical oversight. For results interpretation and protocol routing, see Document 07C — Interpreting Your Results.
| Test | What it measures | Cascade node confirmed | Lab source | Priority | Regression |
|---|---|---|---|---|---|
| Zonulin (serum or stool) | Intestinal tight junction protein. Elevated = active barrier disruption and LPS translocation risk. Most direct measure of leaky gut. | Upstream trigger — LPS entry point. Elevated = gut is active ongoing cascade driver. | Cyrex Array 2; Doctor's Data stool panel | First-line | High |
| LPS-Binding Protein (LBP) | Acute phase protein that binds circulating LPS. Elevated = systemic LPS translocation actively occurring, driving TLR4/NF-κB activation. | Upstream trigger — confirms active LPS-driven NF-κB signaling. Makes gut Layer 1 priority. | LabCorp, Quest (order as LBP) | First-line | High |
| GI-MAP (comprehensive stool) | Pathogenic and commensal bacteria, H. pylori, parasites, fungi, secretory IgA (mucosal defense), calprotectin (gut inflammation), pancreatic elastase (enzyme output). | Upstream trigger + SST node — low elastase confirms SST inhibition of pancreatic secretion. Calprotectin quantifies gut inflammation severity. | Diagnostic Solutions (GI-MAP); Genova GI Effects | First-line | High |
| OAT — gut dysbiosis markers | Arabinose (yeast overgrowth), HPHPA (Clostridia), other dysbiosis metabolites in urine. Captured within the full OAT panel. | Upstream trigger — dysbiosis metabolites confirm gut is driving LPS load and propionate excess. Guides antimicrobial/antifungal sequencing before probiotic protocol. | Great Plains Laboratory OAT (included); Genova Organix | First-line | High |
| Anti-LPS IgG / IgM | Antibodies against LPS — marker of cumulative chronic LPS exposure. Elevated = gut has been leaking LPS for an extended period, not just acutely. | Upstream trigger — confirms chronic vs. acute LPS exposure; guides urgency and depth of gut repair protocol. | Cyrex Array 2; specialty functional labs | Second-line | Conditional |
| Secretory IgA (stool) | Mucosal immune defense layer. Low sIgA = gut immune surveillance compromised; predicts dysbiosis persistence and LPS vulnerability even after antimicrobial treatment. | Upstream trigger — low sIgA predicts failure of probiotic interventions without concurrent sIgA support. Colostrum and saccharomyces boulardii support sIgA restoration. | Included in GI-MAP and Genova GI Effects panels | Second-line | High |
| Test | What it measures | Cascade node confirmed | Lab source | Priority | Regression |
|---|---|---|---|---|---|
| OAT — TCA cycle markers | Succinic, fumaric, malic, and citric acids in urine. Elevated = TCA cycle blockade indicating mitochondrial dysfunction. Also captures lactate and pyruvate metabolites. | Mitochondrial dysfunction node — confirms mitochondrial energy production failure from NAD⁺ depletion and calcium overload secondary to quinolinic acid NMDA overactivation. Two or more elevated TCA markers = mitochondrial channel confirmed. | Great Plains Laboratory OAT (included in full panel); Genova Organix | First-line | High |
| Plasma lactate:pyruvate ratio | Direct measure of mitochondrial respiratory chain function. Ratio >20 at rest = Complex I/III dysfunction with anaerobic glycolysis active. Most specific marker of active respiratory chain failure. | Mitochondrial node — elevated L:P ratio (>25) is the strongest single confirmatory marker of respiratory chain failure from calcium overload and phosphatidylcholine membrane depletion. Distinguishes State 2 metabolic exhaustion from State 1 transcriptional suppression alone. L:P >25 = State 2 biomarker. | LabCorp, Quest standard metabolic panels; specify lactate:pyruvate ratio. Order as fasting, resting sample. | First-line | High — motor regression |
| Plasma / RBC carnitine panel | Free and total carnitine plus acylcarnitine profile. Low free carnitine = impaired beta-oxidation. Acylcarnitine ratios identify specific metabolic blocks in fatty acid oxidation. | PGC-1α / mitochondrial node — carnitine depletion prevents fatty acid entry into the TCA cycle. Acylcarnitine profile guides targeted L-carnitine or acetyl-L-carnitine supplementation. | LabCorp, Quest; request full carnitine panel with acylcarnitine profile. Specify plasma or RBC. | Second-line | Conditional |
| CoQ10 (plasma) | Direct ubiquinol and ubiquinone levels. CoQ10 deficiency is common in mitochondrial dysfunction and in children on certain medications. Guides ubiquinol dose selection. | Mitochondrial / PGC-1α node — low plasma CoQ10 confirms need for ubiquinol (reduced form) supplementation rather than ubiquinone. Dose confirmed by level, not estimated. | LabCorp, Quest; specify CoQ10 plasma (not RBC). Genova NutrEval includes CoQ10. | Second-line | Conditional |
| Test | What it measures | Cascade node confirmed | Lab source | Priority | Regression |
|---|---|---|---|---|---|
| Kynurenine / Tryptophan ratio (K:T) | Direct measure of IDO1 enzyme activity. Elevated K:T = IDO1 activated by inflammation, diverting tryptophan from serotonin synthesis into the kynurenine pathway. Functional NAD⁺ insufficiency confirmed. | IDO1 activation node — most direct single confirmatory test for the Arm A excitotoxic cascade mechanism. Elevated K:T confirms IDO1 is diverting tryptophan into the kynurenine pathway, producing quinolinic acid for NMDA overactivation of SST-14 interneurons and simultaneously depleting NAD⁺ through mitochondrial stress responses. NMN/NR intervention becomes urgent. Elevated K:T also confirms the hypothalamic arm is active: IDO1 diverts tryptophan away from serotonin synthesis, suppressing oxytocin release via the hypothalamic PVN. Launay et al. (2023, Translational Psychiatry, n=271) directly measured K:T ratio, NAD⁺, and plasma oxytocin in the same ASD cohort of 271, establishing both arms as measured human consequences. | Mayo Clinic Labs; Boston Heart Diagnostics; some functional labs. OAT captures downstream kynurenine metabolites as a proxy. | First-line | High — core mechanism |
| OAT — quinolinic / kynurenic ratio | Quinolinic acid (neurotoxic NMDA agonist) versus kynurenic acid (neuroprotective NMDA antagonist). High ratio = neurotoxic kynurenine pathway dominance. Quinolinic acid elevation = active oxidative pressure on neurons. | IDO1 / tryptophan hijack node — elevated quinolinic acid confirms active neurotoxic IDO1 pathway. NAC becomes urgent to protect against quinolinate-driven oxidative DNA damage. Elevated quinolinic acid also signals that the hypothalamic arm is active: the same IDO1 activation depleting NAD⁺ also depletes serotonin, suppressing oxytocin release via the PVN. Both the excitotoxic arm (QUIN → NMDA) and the hypothalamic arm (serotonin depletion → oxytocin suppression) are running simultaneously when this ratio is elevated. | Included in Great Plains OAT full panel; Genova Organix includes kynurenine metabolites. | First-line | High — NMDA pressure |
| hsCRP (high-sensitivity C-reactive protein) | Systemic inflammatory load marker. Elevated >1.0 mg/L in absence of acute infection = chronic NF-κB activation driving the inflammatory cascade. Accessible first-line screen. | NF-κB node — elevated hsCRP confirms chronic inflammatory state. Most accessible entry point for detecting active neuroinflammatory drive. Pair with ESR for confirmation. | LabCorp, Quest standard. Inexpensive, widely covered by insurance. Included in many standard panels. | First-line | High |
| Cytokine panel (IL-6, TNF-α, IL-1β) | Pro-inflammatory cytokines from activated M1 microglia and NF-κB-driven immune cells. IL-6 and TNF-α confirm active M1 microglial state; IL-1β confirms NLRP3 inflammasome activation. | NF-κB / M1 microglia node — together confirms active neuroinflammatory signaling. Luteolin + omega-3 move to Layer 1 when both IL-6 and TNF-α are elevated. | LabCorp, Quest (IL-6, TNF-α available standard); specialty labs for IL-1β. Order as fasting sample to reduce noise. | Second-line | High |
| BDNF (serum) | Downstream output of the cAMP/PKA/CREB pathway — the convergence point where NF-κB-mediated CREB suppression (from inflammatory cytokines), adenosine-driven cAMP deficit (from CD26 blockade), and quinolinic acid excitotoxic depletion all converge to suppress BDNF transcription. Chronically low in ASD. Primary treatment response outcome marker. | CREB/BDNF node — low BDNF confirms SST-14 interneuron silencing has reached the output level, with both NF-κB-mediated CREB suppression and adenosine-driven cAMP deficit reducing BDNF gene transcription. Order at baseline; retest at 3 and 6 months. Rising BDNF = cascade responding to treatment. | LabCorp, Quest; specify serum BDNF. Sample handling matters — EDTA tube, centrifuged promptly. Confirm lab protocol before ordering. | Second-line | Urgent baseline |
Regression — the loss of speech, language, motor skills, or social connection that were previously present — requires a specific test combination rather than a single channel panel. Biologically, regression is the observable downstream consequence of SST-14 interneuron silencing reaching the threshold at which previously encoded neural circuits can no longer be maintained. Both the IDO1 excitotoxic arm and the NF-κB transcriptional suppression arm are typically active simultaneously — the regression test battery must probe both rather than defaulting to one channel's panel.
Regression test battery — order together at first evaluation
| Test | What it confirms in regression | Lab source | Priority | |
|---|---|---|---|---|
| BDNF (serum — urgent baseline) | Directly confirms CREB pathway output is suppressed. The most specific single marker that the learning consolidation circuit has failed. Low BDNF at regression onset sets the recovery benchmark — rising BDNF at 3 and 6 months confirms the protocol is restoring pathway function. | LabCorp, Quest; specify serum BDNF. EDTA tube, centrifuge promptly. | First-line | High |
| OAT — kynurenine metabolites + TCA markers | Quinolinic acid elevation confirms IDO1-driven excitotoxic pressure on SST-14 interneurons (NMDA overactivation, calcium overload, ATP depletion). Elevated TCA markers confirm State 2 metabolic exhaustion — especially relevant when motor regression accompanies speech regression. | Great Plains Laboratory OAT full panel. First-morning urine. | First-line | High |
| K:T ratio (kynurenine:tryptophan) | Confirms IDO1 is actively diverting tryptophan — generating quinolinic acid for SST-14 NMDA excitotoxic pressure and depleting NAD⁺ through mitochondrial stress. In regression, this is often the most actionable finding because NMN/NR can be introduced urgently once confirmed. | Mayo Clinic Labs; Boston Heart Diagnostics. | First-line | High |
| Salivary cortisol (4-point) + IGF-1 | Confirms whether the SST/HPA arm is also suppressing CREB independently. Abnormal cortisol pattern + low IGF-1 means both arms are active simultaneously — CREB is being suppressed from two directions at once, the most treatment-resistant configuration. | ZRT Laboratory (saliva); IGF-1 via LabCorp/Quest standard. | First-line | High |
| Zonulin + LBP (if regression followed a gut event or illness) | When regression followed a febrile illness, vaccination, or GI event, active LPS translocation may have been the immune trigger. Zonulin + LBP confirm whether the gut is still an active upstream driver sustaining the cascade after the initial triggering event. | Cyrex Array 2 (Zonulin); LabCorp or Quest (LBP — order as LPS-Binding Protein). | Second-line | High |
| Plasma lactate:pyruvate ratio + carnitine panel | Add when motor regression accompanies speech or social regression. Elevated L:P ratio confirms respiratory chain dysfunction is contributing independently. Motor regression has a stronger association with active mitochondrial dysfunction than speech regression alone. | LabCorp, Quest; specify fasting resting sample for L:P ratio. | Second-line | High — motor regression |
For the cascade interpretation of regression findings and protocol implications, see Document 07C — Regression interpretation. For scripts to use when discussing regression with a practitioner, see Document 07D — Regression script.
| Test | What it measures | Cascade node confirmed | Lab source | Priority | Regression |
|---|---|---|---|---|---|
| Salivary cortisol (4-point diurnal) | Cortisol at waking, midday, afternoon, and evening. Maps the full diurnal rhythm pattern. Flat or inverted curve = HPA axis dysregulated, SST tonically elevated, cAMP/CREB suppressed around the clock. | SST node — HPA dysregulation is the primary driver of chronic SST elevation. Abnormal pattern confirms AC/cAMP/CREB chronically suppressed. Sleep/circadian optimization becomes non-negotiable Layer 1. | ZRT Laboratory (saliva); DUTCH Test (Precision Analytics); Genova Adrenocortex Stress Profile. Not available through standard LabCorp/Quest. | First-line | High |
| IGF-1 (serum) | SST inhibits growth hormone (GH), which drives IGF-1 production. Low IGF-1 for age = functional downstream marker of chronically elevated SST tone. Documented low in a subset of ASD individuals. | SST node — low IGF-1 confirms SST chronically suppressing the GH axis via a second independent pathway. When both cortisol pattern and IGF-1 are abnormal, SST channel is doubly confirmed. | LabCorp, Quest standard serum test. Inexpensive and widely available. Age-reference ranges are essential — use pediatric norms. | First-line | High |
| Fasting insulin / HOMA-IR | Insulin resistance index. Metabolic strain and hyperinsulinemia are direct SST release triggers — the pancreas responds to elevated insulin demand by increasing somatostatin output. HOMA-IR = fasting insulin × glucose / 405. | SST / upstream metabolic node — elevated HOMA-IR confirms a metabolic component driving SST elevation independent of HPA stress. Dietary intervention and insulin sensitization become protocol priorities. | LabCorp, Quest standard. HOMA-IR requires fasting glucose and fasting insulin drawn at the same time. Specify fasting draw. | First-line | Conditional |
| DHEA-S (serum) | Adrenal androgen produced alongside cortisol. Low DHEA-S with abnormal cortisol pattern = HPA exhaustion — the adrenal axis is in a conservation phase after chronic activation. Chronic SST elevation is strongly confirmed. | SST / HPA node — low DHEA-S + flat cortisol curve is the HPA exhaustion signature. Adrenal recovery must precede cAMP infrastructure restoration. Phosphatidylserine 100–200mg evening is the first intervention. | LabCorp, Quest standard serum test. Inexpensive and widely covered. Interpret always in the context of the cortisol diurnal curve — DHEA-S alone is insufficient. | First-line | Conditional |
| Hair cortisol (3-month integrated) | Cortisol incorporates into the hair shaft at approximately 1 cm/month. A 3 cm proximal segment captures the preceding 3 months of integrated HPA output. Distinguishes chronic load from acute or transient HPA activation. | SST / HPA node — hair cortisol quantifies sustained chronic HPA burden sustaining long-term SST elevation. Complements salivary cortisol: rhythm pattern (saliva) + total load (hair). Elevated hair cortisol = chronic load confirmed, not a transient stress response. | ZRT Laboratory (hair); Stratech/Ciatec; Eurofins; some LabCorp specialty labs. Collect 3 cm proximal segment close to scalp. | Second-line | Conditional |
| Glutamate / GABA ratio | Excitatory / inhibitory (E/I) neurotransmitter balance. Elevated glutamate or depressed GABA = E/I imbalance. Correlates with sensory hypersensitivity, anxiety, and repetitive behaviors in ASD. | SST node + connectivity node — E/I imbalance is the synaptic signature of SST interneuron loss and glypican deficiency. Elevated ratio confirms the cascade has reached architectural impact level, not just signaling suppression. | OAT includes glutamate metabolites (most accessible); plasma amino acid panels (LabCorp, Quest); CSF requires lumbar puncture at specialized centers only. | Second-line | Conditional |
| Actigraphy / sleep study (PSG) | Objective sleep architecture measurement. Identifies slow-wave sleep (SWS) deficit — the glymphatic-active sleep phase when neuroinflammatory debris is cleared. SWS <15% of total sleep time = clearance failure. | SST / glymphatic node — SWS deficit confirms glymphatic clearance is impaired; inflammatory debris (quinolinic acid, cytokines) accumulates nightly. Melatonin timing and sleep optimization become non-negotiable before other interventions. | Polysomnography via sleep specialist for clinical PSG. Consumer actigraphy (Oura Ring, Garmin, Apple Watch) for directional monitoring. Clinical PSG required for formal diagnosis of sleep disorder. | First-line | Conditional |
| Test | What it measures | Cascade node confirmed | Lab source | Priority | Regression |
|---|---|---|---|---|---|
| Neural autoantibody panel | Autoantibodies against neural surface proteins including NMDA receptor subunits, GABAergic interneuron proteins, and brain-specific antigens. Positive titres confirm the adaptive immune system has produced antibodies that jam SST-14 interneuron surface receptors — the State 1 surface receptor jamming mechanism. | NF-κB/autoantibody node — confirms State 1 transcriptional suppression mechanism is active. Autoantibody clearance by IMIG/IVIG is the rate-limiting intervention when this is positive. Result directly informs IMIG candidacy assessment. | Cunningham Panel (Moleculera Labs) for anti-neural antibodies; anti-NMDA receptor antibodies via LabCorp/Quest; ARUP Laboratories neural antibody panel. | Second-line | High — State 1 confirmation |
| IgG subclass panel | IgG1, IgG2, IgG3, IgG4 subclass levels. IgG subclass deficiency — particularly IgG2 deficiency — is consistently elevated in ASD immune dysregulation subgroup and predicts IMIG/IVIG candidacy. Combined with low specific polysaccharide antibody titres confirms the immune insufficiency pattern. | Upstream immune node — IgG subclass abnormalities confirm immune dysregulation pattern. A primary indicator for IMIG/IVIG evaluation. Low IgG2 + elevated inflammatory cytokines = core IMIG candidacy signal. | LabCorp, Quest standard panel. Order with specific polysaccharide antibodies (pre- and post-pneumovax) for full immune evaluation. | Second-line | High |
| Plasma homocysteine | Upstream metabolite of the methylation cycle. Elevated homocysteine (>10 μmol/L) with normal B12 and folate concentrations indicates functional B12 insufficiency — adenosine from CD26 blockade is rate-limiting methionine synthase despite adequate B12. The most accessible biomarker for the CD26/methylation failure arm of the cascade. | CD26/methylation node — the laboratory fingerprint of the Arm 2B cascade mechanism. Elevated homocysteine with normal B12/folate = adenosine rate-limiting methionine synthase, not dietary B12 deficiency. SAMe production is reduced. Methylation cycle restoration (hydroxy-B12, folinic acid, magnesium) becomes a priority intervention. | LabCorp, Quest standard. Widely covered by insurance. Include B12, folate, and methylmalonic acid to distinguish functional from dietary deficiency. | First-line | High |
| SAMe:SAH ratio | Direct quantification of methylation cycle capacity. SAMe is the universal methyl donor; SAH is its product and a competitive inhibitor of methyltransferase enzymes. Low SAMe:SAH ratio directly measures the stalled methylation cycle, not just the upstream homocysteine elevation. Confirms active methyltransferase inhibition from SAH accumulation. | Methylation cycle node — SAMe:SAH directly quantifies the stalled cycle. Low ratio confirms active suppression of neurotransmitter regulation (COMT), immune cell switching, DNA methylation, and phosphatidylcholine synthesis simultaneously. Most specific methylation cycle marker; complements homocysteine measurement. | Specialty metabolic labs: Genova Diagnostics, Doctor's Data, LabCorp specialty testing. Not routinely available at standard draw sites — may require advance ordering. | Second-line | Medium |
| DPP-IV / CD26 activity (serum) | Enzymatic activity of dipeptidyl peptidase IV (DPP-IV / CD26) in serum. Reduced activity confirms CD26 receptor blockade — the docking site for adenosine deaminase has been occupied by casomorphin, gliadorphin, streptokinase, or mercury, preventing adenosine clearance and initiating the methylation failure arm. Documented at reduced levels in ASD populations by multiple independent groups. | CD26/adenosine node — reduced DPP-IV activity confirms Arm 2B mechanism is operative. Clinically actionable: identifies children for whom dietary elimination (casein/gluten) has the highest mechanistic rationale, and for whom streptococcal infection management and mercury exposure history are most relevant upstream drivers. | Specialty functional labs: AAL Reference Laboratories, Metametrix (now Genova), some European functional labs. Not yet available through LabCorp/Quest standard panels as of 2025 — may require specialty ordering. | Second-line | Medium |
| Urinary opioid peptides (casomorphin / gliadorphin) | Casomorphin (from incompletely digested casein) and gliadorphin (from incompletely digested gluten) detectable in urine. Confirms that intact opioid peptide fragments have crossed the gut barrier and entered systemic circulation — directly demonstrating active pepsin failure and the CD26 blockade upstream trigger. | Pepsin failure / CD26 blockade node — positive result confirms the upstream dietary peptide mechanism is operative. Clinically defines the population for whom casein- and gluten-free dietary elimination has highest mechanistic rationale. Explains food selectivity as a biological drive, not a behavioral preference. Opioid withdrawal responses during dietary elimination are predictable when this test is positive. | Great Plains Laboratory (peptide panel); AAL Reference Laboratories; Genova Diagnostics. First-morning urine collection, frozen shipping required. | Second-line | Medium |
| Plasma NAD⁺ | Direct measurement of intracellular NAD⁺ availability. NAD⁺ is the energy substrate required for SST-14 interneuron tonic high-frequency firing. Depleted by chronic IDO1 overactivation consuming NAD⁺ through the kynurenine pathway faster than it can be replaced, and by CD38-mediated NAD⁺ consumption during immune activation. The State 2 metabolic exhaustion primary marker. | NAD⁺ depletion node — low plasma NAD⁺ confirms State 2 metabolic exhaustion mechanism. NMN or NR supplementation is the targeted response. NAD⁺ level also calibrates the urgency: severely depleted (<20 μM) indicates SST-14 interneurons are energy-limited and cannot maintain tonic firing even if transcriptional suppression were removed. State 2 treatment sequence is determined by this finding. | Specialty labs: Jinfiniti Precision Medicine (NAD⁺ test from finger prick); some functional medicine labs. Not available through standard LabCorp/Quest panels. Finger-prick at-home collection available through Jinfiniti. | Second-line | High — State 2 |
| Folate receptor alpha antibodies (FRAA) | Autoantibodies against the folate receptor alpha that block methylfolate transport into the brain and CSF, independently of dietary folate status. Present in a significant subset of ASD children. Creates a dual methylation vulnerability when combined with cascade-driven adenosine accumulation: methionine synthase rate-limited from two independent directions simultaneously. | Dual methylation vulnerability node — FRAA positive + elevated homocysteine identifies the subgroup for whom folinic acid (not folic acid) is specifically indicated. The Frye et al. folinic acid RCT (Mol Psychiatry 2018) — the single published ASD intervention with consistent positive results — targeted precisely this mechanism. FRAA positive result changes the methylation intervention from general support to specific folinic acid protocol. | Iliad Neurosciences (formerly Ramaekers lab) — the only commercially available FRAA test as of 2025. Requires specialist ordering through Iliad's clinician portal. | Second-line | High if methylation not responding |
| Test | What it measures | Cascade node confirmed | Lab source | Priority | Regression |
|---|---|---|---|---|---|
| GPL-TOX (toxic organic chemicals) | Urine screen for 172+ environmental chemicals including glyphosate, organophosphates (pesticides), phthalates (plastics), benzene, styrene, and MTBE. Captures both agricultural and urban chemical exposure. | Upstream trigger — environmental channel. Positive findings confirm ongoing toxin-driven oxidative load contributing to IDO1 activation, gut microbiome disruption, and mitochondrial impairment simultaneously. | Great Plains Laboratory (GPL-TOX urine panel). Direct-to-consumer ordering available in most states. | First-line | Conditional |
| Glyphosate (urine, specific) | Quantitative glyphosate measurement. Glyphosate disrupts gut microbiome by inhibiting the shikimate pathway in bacteria, depletes tryptophan precursors, and is epidemiologically associated with increased ASD risk. | Upstream trigger — glyphosate depletes Lactobacillus and Bifidobacterium, compounding IDO1 pathway dysregulation and gut barrier vulnerability. Positive = gut repair more urgent, dietary glyphosate reduction essential. | Great Plains Laboratory (included in GPL-TOX); The Detox Project; HRI Labs. Can be ordered standalone or as part of GPL-TOX. | First-line | Conditional |
| RBC heavy metals (Hg, Pb, As, Cd) | Red blood cell metal levels reflecting chronic tissue exposure rather than recent acute exposure (serum measures only recent). Mercury disrupts mitochondria; lead elevates inflammatory cytokines; arsenic directly impairs mitochondrial enzyme function. | Upstream trigger + FOXO node — metal burden drives oxidative stress, GSH depletion, and mitochondrial impairment from a second independent direction. Must specify RBC — serum metals miss chronic tissue burden. | Doctor's Data RBC Elements panel; LabCorp metals panel with RBC specification. Must specify RBC not serum when ordering. | First-line | Conditional |
| 8-OHdG (urine) | 8-hydroxy-2-deoxyguanosine — a DNA oxidative damage marker produced when reactive oxygen species modify guanine. Elevated in ASD and correlates with behavioral severity. Confirms active oxidative DNA damage. | FOXO / antioxidant node — elevated 8-OHdG confirms ROS burden has reached genomic level. NAC + sulforaphane are prioritized to restore antioxidant enzyme capacity. Retest at 3 months to confirm response. | Great Plains OAT includes 8-OHdG (most accessible); standalone via LabCorp. Also included in some comprehensive oxidative stress panels. | Second-line | Lower priority |
| Glutathione GSH:GSSG ratio | Ratio of reduced (active) to oxidized glutathione. Low ratio = antioxidant system overwhelmed and FOXO axis depleted. Quantifies antioxidant depletion depth more precisely than 8-OHdG alone. | FOXO / antioxidant node — low GSH:GSSG is the primary indicator for urgency of NAC intervention. Severely low ratio (<80% GSH) may indicate need for direct liposomal glutathione in addition to NAC. | Doctor's Data; Genova Diagnostics; some specialty labs. Not available through standard LabCorp/Quest — requires functional lab ordering. | Second-line | Lower priority |
| Test | What it measures | Cascade nodes covered | Lab source | Priority | Regression |
|---|---|---|---|---|---|
| OAT — full panel | Single first-morning urine collection covering: mitochondrial TCA cycle markers, kynurenine/IDO1 pathway metabolites, gut dysbiosis metabolites (arabinose, HPHPA), oxidative stress markers (8-OHdG), neurotransmitter metabolites, and B-vitamin functional status. | Covers IDO1, mitochondrial, gut/LPS, FOXO/antioxidant, and CREB nodes in a single collection. Highest-yield single test in the entire testing suite for initial channel mapping. | Great Plains Laboratory OAT (most comprehensive version). Genova Organix as alternative. First-morning urine; no vitamin B supplementation 48 hours prior. | First-line | High — start here |
| Intracellular micronutrient panel | Intracellular (not serum) levels of B vitamins, zinc, magnesium, selenium, CoQ10, vitamin D, and antioxidants. Co-factor deficiencies limit the efficacy of every cascade node intervention — supplements underperform without adequate cofactors. | All nodes — must be identified before building the intervention protocol. B6 and magnesium have the strongest ASD-specific RCT data. Zinc is essential for metallothionein and gut barrier function. Selenium is required for glutathione peroxidase. | Spectracell Micronutrient Panel; Genova NutrEval; LabCorp ION Panel. Serum nutrient levels are insufficient — intracellular or functional testing required for meaningful results. | First-line | Conditional |
| Vitamin D (25-OH-D) | VDR (vitamin D receptor) modulates NF-κB transcription. Deficiency is near-universal in ASD, correlates with severity, and permits unchecked NF-κB activation. Target range 50–70 ng/mL. | NF-κB / microglial node — deficiency amplifies the inflammatory arm of the cascade from a separate direction. Inexpensive to test, easy to correct. Order for every individual regardless of other findings. | LabCorp, Quest standard. Inexpensive, widely covered by insurance. Order as 25-OH-D (calcidiol), not 1,25-OH-D (calcitriol). | First-line | High |
| CBC + CMP | Complete blood count (CBC) and comprehensive metabolic panel (CMP). Screens for anemia, liver and kidney function, glucose and insulin indicators, and inflammatory markers. Universal safety baseline before initiating any supplement protocol. | Multi-node baseline — identifies systemic constraints on detoxification, nutrient delivery, and metabolism that limit protocol efficacy. Abnormal LFTs or creatinine require clinical review before hepatically-cleared supplements. | LabCorp, Quest standard. Covered by virtually all insurance. Available through primary care without specialist referral. | First-line | High |
| ApoE genotyping | ApoE4 allele impairs glymphatic clearance of neuroinflammatory debris, Aβ, and tau during sleep. ApoE4 carriers have 3–4× higher risk of neuroinflammatory debris accumulation for any given level of sleep disruption. | Glymphatic / sleep node — ApoE4 status identifies individuals where sleep disruption has compounding neuroinflammatory consequences. Melatonin timing and circadian repair become non-negotiable Layer 1 in ApoE4 carriers. Once-in-lifetime result. | LabCorp, Quest standard genetic test. Single blood draw. 23andMe raw data can be parsed for ApoE status via Promethease or similar. | Second-line | Lower priority |
The following sequence is the recommended ordering framework for families working through the full testing bridge. Each stage builds on the previous — second-line tests are only meaningful after first-line results establish which channels are active. The sequence prevents over-testing before direction is established and under-testing once a channel is confirmed.
| Stage | Tests | What you learn | Action threshold |
|---|---|---|---|
| 1 | Map active channels OAT (full panel) · Intracellular micronutrient panel · Vitamin D (25-OH-D) · CBC + CMP · hsCRP | Active channels across gut, mitochondria, IDO1/kynurenine, and oxidative stress identified in a single round. Co-factor depletions limiting all subsequent interventions identified. Baseline inflammatory status and safety markers captured. | OAT abnormalities route to channel-specific Stage 2 tests. Vitamin D <40 ng/mL: supplement immediately regardless of other findings. CMP abnormalities: clinical review before advancing protocol. hsCRP elevated: anti-inflammatory layer prioritized. |
| 2 | Confirm channel dominance K:T ratio · Salivary cortisol (4-point) · Hair cortisol (3-month) · IGF-1 · DHEA-S · Fasting insulin / HOMA-IR · Zonulin + LBP (if OAT gut markers elevated) | Primary cascade driver identified and quantified. SST/HPA load assessed from two angles — rhythm (saliva) and chronic burden (hair). Gut as active vs. resolved ongoing trigger clarified. Protocol layer reordering determined. | K:T elevated: NMN/NR to Layer 1 urgently. Abnormal cortisol curve: sleep/circadian as Layer 1. Elevated hair cortisol: chronic HPA load confirmed. Low IGF-1 + abnormal cortisol: SST doubly confirmed. Low DHEA-S + flat cortisol: HPA exhaustion pattern — adrenal recovery before cAMP infrastructure restoration. Elevated Zonulin + LBP: gut repair before all other layers. |
| 3 | Quantify severity Cytokine panel (IL-6, TNF-α) · GSH:GSSG ratio · BDNF (serum baseline) · GI-MAP stool (if gut channel confirmed active) | Neuroinflammation severity and M1 microglial activation depth mapped. Antioxidant depletion depth quantified. Learning circuit impairment baseline established via BDNF. Gut microbiome composition and pathogen load detailed for targeted antimicrobial protocol. | Cytokines both elevated: luteolin + omega-3 move to Layer 1; curcumin added as adjunct. Low GSH:GSSG: NAC becomes urgent; liposomal glutathione considered if severely low. Low BDNF: establishes baseline — retest at 3 and 6 months. GI-MAP calprotectin very high: gastroenterology review before probiotic protocol. |
| 4 | Environmental burden GPL-TOX + glyphosate (urine) · RBC heavy metals · 8-OHdG (if not captured in OAT) · Carnitine panel (if mito channel confirmed) | Environmental toxin contribution to the cascade quantified. Metal burden and its impact on mitochondrial function and GSH depletion identified. Mitochondrial fatty acid oxidation capacity assessed when TCA markers were elevated in Stage 1. | GPL-TOX positives: detox support (NAC, sulforaphane, glycine) + dietary exposure reduction. Metals elevated: NAC/glutathione urgently; physician-supervised chelation consultation if Hg or Pb significantly elevated. Low carnitine: L-carnitine 500–1000mg with meals. 8-OHdG elevated: NAC + sulforaphane urgently. |
Many of the tests in this reference are not available through standard pediatric primary care. Understanding where each type of test lives — and how to access it — is essential to navigating the testing bridge without unnecessary friction.
Most Stage 1 baseline tests and many Stage 2 confirmatory tests are available through LabCorp and Quest via a physician order or direct-to-consumer. This includes CBC, CMP, hsCRP, ESR, vitamin D, IGF-1, DHEA-S, fasting insulin, cytokines (IL-6, TNF-α), RBC metals, serum BDNF, and CoQ10.
The OAT, GI-MAP, GPL-TOX, salivary cortisol, hair cortisol, GSH:GSSG ratio, and intracellular micronutrient panel require specialty labs not connected to LabCorp/Quest.
If your current pediatrician is unfamiliar with functional panels, these practitioner types typically order them routinely:
Insurance coverage varies significantly by plan and by test type.