Skip to main content
. 2017 Dec 8;17:836. doi: 10.1186/s12885-017-3820-5

Table 1.

Study and population characteristics

Study Participant selection and disease prevalence a Participant characteristics Reference standard Microdissection MSI Panel MSI thresholds
Population-based and age-limited single-gate studies
 Barnetson, 2006 Diagnosed <55yrs of age, consecutive recruitment
Disease prevalence 8.5% (95% CI 5.8 to 11.9)
Number: For all participants: Germ-line DNA obtained from blood leukocytes analysed for MLH1, MSH2, and MSH6 mutations. dHPLC analysis was used for MSH2 and MLH1. Variants noted on chromatography were then sequenced.
Mutations were confirmed by re-amplification of an independent sample of DNA and resequencing in both directions. MLH1 and MSH2 were assessed for deletions by MLPA, with products separated on a genetic analyser.
10μm tumour sections; microdissection performed on purified tumour DNA, and control DNA from blood or normal tissue in the section Bethesda/NCI panel MSI-H: >1 marker
MSI-L: 1 marker
MSS: 0 markers
Recruited 1259
Receiving RS 870
Receiving MSI 352
Gender:
Male 53.1%
Female 46.9%
Age (in years):
Non-carrier 48.2 (±6.0)
Carrier 42.7 (±7.7)
MLH1 38.5 (±8.4)
MSH2 43.8 (±6.1)
MSH6 49.0 (±3.9)
Clinical criteria:
AMS II 4.0%
RBG 64.0%
 Poynter, 2008b Recruitment through population-based cancer registries (population-based sample), selection process unclearc
Disease prevalence 9.2% (95% CI 7.1 to 11.8)
Number: For all MSI-H or MSI-L probands and in a random sample of 300 MSS population-based probands: Mutations in MSH2 and MLH1 were detected using a combined approach of dHPLC/direct sequencing and MLPA. Direct sequencing was used to detect MSH6 mutations in cases with absent IHC staining of MSH6. NR BAT25, BAT26, D5S346, D17S250, BAT40, MYCL, ACTC,
Dl 8S55, D1OS197, BAT34C4
MSI-H: ≥30%
MSI-L: >0% and <30%
MSS: 0%
Recruited 1061
Receiving RS 726
Receiving MSI 1061
Gender: NCR
Age (in years): NCR
Clinical criteria: NCR
 Southey, 2005 Diagnosed <45yrs of age, random recruitment
Disease prevalence 30.5% (95% CI 19.2 to 43.9)
Number: For all MSI-H or MSI-L probands, those that lacked expression of at least one MMR protein and a random sample of 23 patients selected from those who had tumours that were MS stable and did not lack expression of any MMR protein: MLH1, MSH2, MSH6, and PMS2 genes were screened for germline mutations using sequencing approaches or dHPLC. Confirmation of putative mutations was sought using an independent polymerase chain reaction for direct automated sequencing. MLPA was used to detect large genomic alterations in MLH1 and MSH2 on samples from 10 patients who had tumours lacking at least one MMR protein expression and for which no previous mutation had been identified by sequencing. 5μm tumour sections; microdissection performed on invasive tumour cells from paraffin-embedded archival tumour tissue stained with 1% methyl-green, and normal cells from colonic or lymph node tissue/DNA extracted from peripheral blood lymphocytes BAT25, BAT26, D2S123, D5S346, D17S250, BAT40, MYB, TGFRII, IGFIIR,
BAX
MSI-H: >5 markers
MSI-L: 2-5 markers
MSS: <2 markers
Recruited 131
Receiving RS 59
Receiving MSI 105
Gender:
Male 62.7%
Female 37.3%
Age (in years):
37.1 (range 24 to 42)
Clinical criteria:
AMS II 9.2%
RGB NR
High-risk, single-gate studies
 Caldes, 2004 HNPCC families selected through a clinic for familial cancer, selection process unclear
Disease prevalence 58.6% (95% CI 44.9 to 71.4)
Number: For all participants: Genomic DNA was isolated from peripheral blood lymphocytes was analysed for MLH1, MSH2 and MSH6. DNA was amplified using PCR and all amplicons were subjected to DGGE or cycle sequencing. The MSI-H cases that were negative for mutations were analysed for genomic deletions in MLH1 and MSH2 by Southern Blotting. 10μm tumour sections; microdissection performed on H&E stained slides with demarked areas containing cancer cells, and corresponding areas on unmarked slides Bethesda/NCI panel MSI-H: >1 marker, or 1 marker if BAT26
MSI-L: Not used
MSS: 0 markers
Recruited 58
Receiving RS 58
Receiving MSI 58
Gender: NR
Age (in years): NR
Clinical criteria: NR
 Mueller, 2009 ‘Suspected Lynch syndrome’ participants who met Amsterdam criteria, modified Amsterdam criteria, were ‘HNPCC-like’ or met Bethesda criteria, selection process unclear
Disease prevalence 58.3% (95% CI 43.2 to 72.4)
Number: For all participants: Sequencing and MLPA. Limited details provided. NR 5 and 10 panel markers, no further details provided NR
Recruited 48
Receiving RS 48
Receiving MSI 48
Gender: NR
Age (in years): NR
Clinical criteria: NR
 Overbeek, 2007 Families history that fulfilled one of the following criteria: 1) Amsterdam II criteria 2) Bethesda guidelines 3) a history very close to the Bethesda guidelines, selection process unclear
Disease prevalence NC
Number: For all participants: Mutation analysis of MLH1, PMS2, MSH2, and MSH6 was performed in DNA from peripheral blood lymphocytes by a combination of either single-strand conformation polymorphism analysis or DGGE and direct sequence analysis.
For the detection of large deletions and duplications in MLH1, MSH2, MSH6, and PMS2, MLPA was used. All deletions and duplications were confirmed by Southern blot analysis or with a specific PCR.
NR BAT25, BAT26, D2S123, D5S346, D17S250 (BAT40 was also added to the standard set of markers but it is unclear for which participants) Tumours categorised as positive (>2 Bethesda markers) or negative
Recruited NR
Receiving RS NR
Receiving MSI NR
Gender: NR
Age (in years): 40.7 (range 29 to 51)
Clinical criteria: NR
 Poynter, 2008b Recruitment through high-risk clinics (clinic-based sample), selection process unclearc
Disease prevalence 30.9% (95% CI 23.7 to 38.9)
Number: For all participants: Mutations in MSH2 and MLH1 were detected using a combined approach of dHPLC/direct sequencing and MLPA.
Direct sequencing was used to detect MSH6 mutations in cases with absent IHC staining of MSH6.
NR BAT25, BAT26, D5S346, D17S250, BAT40, MYCL, ACTC, Dl 8S55, D1OS197, BAT34C4 MSI-H: ≥30%
MSI-L: >0% and <30%
MSS: 0%
Recruited 172
Receiving RS 152
Receiving MSI 172
Gender: NR
Age (in years): NR
Clinical criteria: NR
 Shia, 2005 Family history that fulfilled one of the following criteria: I ) Amsterdam I or II criteria 2) a set of relaxed AC three or more colorectal cancers among the first and second-degree relatives of a family that we referred to as ''HNPCC-like,'' and 3) Bethesda criteria, selection process unclear
Disease prevalence 49.2 (95% CI 36.1 to 62.3)
Number: For all participants: Each of the exons of MLH1, MSH2 and MSH6 was amplified by PCR, and heteroduplex analyses were performed using dHPLC. DNA fragments that displayed an abnormal chromatogram were sequenced directly.
Cases with tumours that exhibited MSI but in which a point mutation was not detected were analysed for large deletions in MLH1 and MSH2 using a procedure based on the multiplex PCR of short fluorescent fragments.
Microdissection performed on DNA from paraffin-embedded tissue blocks. No further details reported BAT25, BAT26, D2S123, D17S250, BAT40, PAX6, MYCL1 Tumours categorised as positive (≥30%) or negative
Recruited 83
Receiving RS 83
Receiving MSI Unclear d
Gender:
Male 43.6%
Female 56.3%
Age (in years):
50 (range 23 to 78)
Clinical criteria:
AMS II 38.2%
 RBG 8.2%
Reference standard positive study
 Hendriks, 2003 Germline mutation in MLH1, MSH2 or MSH6, selection process unclear
Disease prevalence 84.8% (95% CI 68.1 to 94.9)e
Number: For all participants: DGGE or Southern blotting. Limited details provided Microdissection not specifically reported, paired tumour and normal tissue DNA samples were used BAT25, BAT26, D2S123, D5S346, D17S250, BAT40, MSH3 and MSH6 MSI-H: >1 Bethesda markers
MSI-L: 1 Bethesda marker
MSS: 0 Bethesda markers
Recruited 45
Receiving RS 45
Receiving MSI 33
Gender:
Male 35.6%
Female 40.0%
Age (in years):
MLH1 48 (range 29 to 90)
MSH2 40 (range 23 to 61)
MSH6 62 (range 26 to 84)
Clinical criteria: NR

NC not calculable, total number of participants with CRC not reported, NR Not reported, NCR Not clearly reported; aDisease prevalence calculations were based on participants who received both the index test and the reference standard and for whom data were reported. The disease prevalence may, therefore not be an accurate representation of the prevalence in the recruited population, and this is more likely in studies that did not aim to assess the reference standard in all recruited participants; for example, two study samples (the population based sample in Poynter, 2008, and Southey, 2005) did not perform the reference standard in all participants with an MSS result. bPoynter (2008) reports data from two distinct samples, a population-based sample and a high-risk sample; cAlthough Poynter (2008) reports that ‘some centres recruited all incident cases of CRC while others over­ sampled cases with a family history or early age of onset’ it is not clear whether this applies to the high-risk sample alone or in part to the high-risk sample and in part to the population based sample; d MSI data are available for 61 participants but it is unclear how many received the test; eAlthough only reference standard positives were recruited, this included those with an unclassified variant, so when those unclassified variants were considered to be reference standard negatives disease prevalence is 84.8%