Cited by 271Sperm and Testicular FunctionMale Reproductive Health StudiesReproductive Health and Technologies

The DOI matches its Crossref record; 63 of the 63 references it cites were found in a registry or an index; 21 of the 45 sources its claims cite were checked for support.

The Sixth Edition of the WHO Manual for Human Semen Analysis: A Critical Review and SWOT Analysis

Open access

Abstract

Semen analysis is the cornerstone of male fertility evaluation with WHO guidelines providing the basis for procedural standardization and reference values worldwide. The first WHO manual was published in 1980, and five editions have been subsequently released over the last four decades. The 6th Edition was published in July 2021. In this review, we identify the key changes of this 6th Edition. Additionally, we evaluate the utility of this 6th Edition in clinical practice using SWOT (strengths, weaknesses, opportunities, and threats) analysis. This new Edition has made the analysis of basic semen parameters more robust, taking into account the criticisms and grey areas of the previous editions. The tests assessing sperm DNA fragmentation and seminal oxidative stress are well-described. The main novelty is that this latest edition abandons the notion of reference thresholds, suggesting instead to replace them with “decision limits”. While this seems attractive, no decision limits are proposed for either basic semen parameters, or for extended or advanced parameters. This critical review of the 6th Edition of the WHO laboratory manual combined with a SWOT analysis summarizes the changes and novelties present in this new Edition and provides an in-depth analysis that could help its global use in the coming years.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

This critical review summarizes key changes in the 6th Edition of the WHO Manual for Human Semen Analysis and evaluates its clinical utility using SWOT analysis. It finds the 6th Edition improves basic semen analysis and adds SDF/oxidative stress testing, but abandons reference thresholds without providing replacement decision limits.

Key findings

  • The 6th Edition, published July 2021, abandons reference thresholds and suggests replacing them with “decision limits,” yet no decision limits are proposed for basic, extended, or advanced semen parameters.
  • Its reference data combine the 5th Edition and new data from 2010–2020, totaling 3589 fertile men (1800 from the 5th Edition and 1789 new subjects); the 5th centiles are not significantly different from the 2010 WHO 5th Edition.
  • SDF and seminal oxidative stress tests are well-described, but SDF has no thresholds and each laboratory is recommended to develop its own reference range; clinical indications and utility are not discussed.
  • Basic semen analysis changes include re-adopting grade a/b progressive motility, recommending vitality assessment when total sperm motility is below 40%, requiring 200 spermatozoa per replicate, and more precise assessment of concentrations <2 × 10^6/mL.

Methods

  • Design: narrative critical review of the WHO 6th Edition combined with SWOT (strengths, weaknesses, opportunities, threats) analysis.
  • Data/sample: comparison of the 5th and 6th Editions, including the 6th Edition’s combined semen data from 3589 fertile men and discussion of extended/advanced tests, sperm preparation, and cryopreservation.
  • Main analysis: qualitative evaluation of methodological changes, clinical utility, and external/internal factors affecting global use.

Limitations

  • The paper states that some geographical areas, such as South America and Sub-Saharan Africa, remain under-represented in the 6th Edition.
  • The paper states that the 6th Edition lacks decision limits, SDF thresholds, clinical guidance for SDF/genetic tests, and accepted cut-off values for seminal oxidative stress.
  • Evident from the text: this is a narrative critical review and SWOT analysis without a stated systematic search strategy or primary data collection; the SWOT assessment is qualitative.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.3390/life11121368

Cites

63 of 63 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Figures

Figure 1 SWOT (“strengths” (S), “weaknesses” (W), “opportunities” (O) and “threats” (T)) analysis of the 6th Edition of the WHO manual for human semen analysis. SA: semen analysis. SDF: sperm DNA fragmentation. FISH: fluorescent in situ hybridization. OS: oxidative stress.
Figure 1 SWOT (“strengths” (S), “weaknesses” (W), “opportunities” (O) and “threats” (T)) analysis of the 6th Edition of the WHO manual for human semen analysis. SA: semen analysis. SDF: sperm DNA fragmentation. FISH: fluorescent in situ hybridization. OS: oxidative stress.CC BY · PubMed Central

Verification

Claims the paper makes

38 No basis given · 19 Own interpretation · 12 Not checked yet · 7 Partly supported · 3 Supported · 3 No full text of the sources · 2 Supported so far · 2 Sources not found · accuracy not yet measured

Verify all 13 references — free account

Show all 20 claims

  • Probably not a substantial claim. 59
  • It may state two separate claims as one. 28
66 of 66
  • The paper relies on this without support.

    Clinicians and laboratories rely on WHO reference values in clinical practice.

    “To rely on other reference values, clinicians will need to look for other sources in the literature.”
    “Therefore, a possibility remains that clinicians will continue to use the 5th centile values, which were designed in the 5th Edition to compare fertile and infertile men with the criterion of time to pregnancy ≤12 months.”
  • The paper relies on this without support.

    Professional societies have not yet fully endorsed oxidative stress (OS) as a diagnostic tool.

    “Despite more evidence being published recently [55], professional societies have not yet fully endorsed OS as a diagnostic tool.”
  • The paper relies on this without support.

    There are a number of different methodologies for the determination of seminal OS with different results described.

    “However, as there are a number of different methodologies for the determination of seminal OS with different results described,”
  • The paper relies on this without support.

    Seminal plasma is regarded as the human body fluid with the highest amount of antioxidants.

    “which is regarded as the human body fluid with the highest amount of antioxidants.”
  • The paper relies on this without support.

    The MACS technique is not approved for human in vivo use in the United States due to the lack of safety studies on paramagnetic beads in the animal model.

    “Furthermore, the MACS technique is not approved for human in vivo use in the United States (and maybe in some other advanced countries as well) due to the lack of safety studies on paramagnetic beads (used in MACS columns to trap sperm with high SDF) in the animal model.”
5 of 5

Citations

Cites

Show all 63 references

Research works printed but not linked: 3

  1. World Health Organization (2010). WHO Laboratory Manual for the Examination and Processing of Human Semen, WHO Press. [5th ed.].
  2. World Health Organization (1999). WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction, Cambridge University Press. [4th ed.].
  3. Morris, 2019, Prediction of successful ICSI cycles by Oxidation-reduction potential (ORP) and sperm DNA fragmentation (SDF) analysis. A Prospective study, Hum. Reprod., 34, i8

Cited by

Sorted by citations

Items 201–271 of 271

Metadata

Authors
Boitrelle, F., Shah, R., Saleh, R., Henkel, R., Kandil, H., Chung, E., Vogiatzi, P., Zini, A., Arafa, M., & Agarwal, A.
Affiliations
  • Université de Versailles Saint-Quentin-en-Yvelines (FR)
  • Lilavati Hospital & Research Centre (IN)
  • Sohag University (EG)
  • Cleveland Clinic (US)
  • Fakih IVF Fertility Center, Abu Dhabi 31452, United Arab Emirates
  • The University of Queensland (AU)
  • Andromed Health & Reproduction, Fertility Diagnostics Laboratory, Maroussi, 15126 Athens, Greece
  • McGill University (CA)
Corresponding author
Agarwal, Ashok
Venue
Life
Publisher
MDPI AG
Published
2021-12-09
Volume
11
Issue
12
Pages
1368
ISSN
2075-1729 (ISSN-L)
OpenAlex
W4200457040
Cited by
271
References
63
Citations by year
2026: 40 · 2025: 53 · 2024: 96 · 2023: 59 · 2022: 23
FWCI
31.25
Type
journal article
Language
en
Open access
Locations
  • Life (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
  • HAL (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • UWC Research Repository (University of the Western Cape) (submittedVersion)
  • Spiral (Imperial College London) (submittedVersion, cc-by)
Topics
  • Health Sciences › Medicine › Reproductive Medicine › Sperm and Testicular Function
  • Health Sciences › Medicine › Urology › Male Reproductive Health Studies
  • Health Sciences › Medicine › Reproductive Medicine › Reproductive Health and Technologies
Keywords
semen analysis, male fertility, semen parameters, reference thresholds, decision limits, sperm DNA fragmentation, seminal oxidative stress, semen quality assessment, clinical practice

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