Role of one-step nucleic acid amplification in colorectal cancer lymph node metastases detection

Table 1 Characteristics of one-step nucleic acid amplification studies in colorectal cancer patients

Ref.	Nation	Type of study	Patients number (sample number)	Tumor type	Purpose of the OSNA analysis
Croner et al[11], 2010	Germany	Prospective study	184 (184)	Colorectal	Diagnosis of LN metastasis
Yamamoto et al[16], 2011	Japan	Prospective multicenter study	85 (385)	Colorectal	Diagnosis of LN metastasis
Güller et al[12], 2012	Switzerland	Prospective study	22 (313)	Colon	Diagnosis of LN metastasis
Yamamoto et al[13], 2013	Japan	Not shown	30 (66)	Colorectal	Identification of CK19
Croner et al[14], 2014	Germany	Prospective multicenter study	103 (1594)	Colon	Pathologically nodenegative CC
Vogelaar et al[15], 2014	Switzerland	Prospective multicenter study	128 (325)	Colon	Diagnosis of SLN metastasis
Yamamoto et al[17], 2016	Japan	Prospective multicenter study	204 (1925)	Colorectal	Diagnosis of LN metastasis
Aldecoa et al[133], 2016	Spain	Prospective multicenter study	149 (1940)	Colorectal	Correlation between TTL and tumor’s characteristics
Rakislova et al[125], 2017	Spain	Observational study	188 (3206)	Colon	Diagnosis of pooled LN metastasis
Miyake et al[193], 2017	Japan	Prospective study	25 (306)	Rectum	Indication of LPLN dissection
Marhic et al[184], 2017	France	Prospective study	17	Colon	Diagnosis of SLN metastasis
Colling et al[129], 2-17	United Kingdom	Prospective study	19 (82)	Colorectal	Diagnosis of LN metastasis
Aldecoa et al[124], 2017	Spain	Prospective study	71 (936)	Colon	OSNA with endoscopic tattooing
Yeung et al[130], 2018	United Kingdom	Prospective study	16 (78)	Colorectal	OSNA with ICG detection
Brito et al[126], 2018	Portugal	Prospective multicenter study	59 (753)	Colon	Pathologically node negative CRC
Esposito et al[127], 2019	Italy	Prospective study	34 (51)	Colorectal	Diagnosis of SLN metastasis
Diaz-Mercedes et al[109], 2019	Spain	Prospective study	17 (980)	Colorectal	Budget impact analysis
Itabashi et al[18], 2020	Japan	Prospective multicenter study	195	Colorectal	Prognostic value of the OSNA assay for pStage IICRC patients
Archilla et al[145], 2021	Spain	Retrospective multicenter study	342 (5931)	Colorectal	Correlation between the TTL with patient outcome
Weixler et al[147], 2021	Netherlands, Germany, Switzerland	Retrospective multicenter study	87	Colon	Prognostic value of OSNA
Tani et al[33], 2021	Japan	Prospective multicenter study	92	Colon	Diagnosis of pooled LN metastasis
Numata et al[188], 2021	Japan	Prospective study	34	Rectum	Indication of LPLN dissection

CRC: Colorectal cancer; ICG: Indocyanine green; LPLN: Lateral pelvic lymph node; OSNA: One-step nucleic acid amplification; SLN: Sentinel lymph node; TTL: Total tumor load.

Table 2 A comparison of the diagnostic accuracy of the one-step nucleic acid amplification assay in colorectal cancer patients

Ref.	Pathological evaluation	IHC	LN number inspected by OSNA, mean	Sensitivity, %	Specificity, %	Concordance, %	PPV, %	NPV, %
Croner et al[11]	Multi-slice	CK19	1.0	92.5	96.5	93.6	88.1	97.9
Yamamoto et al[16]	Multi-slice	CK19	4.5	95.2	97.7	97.1	91.9	98.7
Güller et al[12]	Multi-slice	CK19	14.2	94.5	97.6	97.1	89.7	98.8
Yamamoto et al[17]	Single-slice	None	9.4	86.2	96.5	95.7	66.5	98.8
Colling et al[129]	Single-slice	None	4.3	92.9	97.1	96.3	86.7	98.5
Yeung et al[130]	Single-slice	None	4.9	100	98.4	98.7	94.1	100
Esposito et al[127]	Multi-slice	None	1.5	69.2	100	88.2	100	84.0
Rakislova et al[125]	Not shown	None	20.5 (pOSNA)	88.9	79.2	80.2	33.3	98.4
Vogelaar et al[15]	Multi-slice	Anti pan-cytokeratin	15.3	51.6	84.1	67.7	76.7	63.1
Miyake et al[193]	Single-slice	CEA	11	100	86	88	57	100
Marhic et al[184]	Not shown	None	Not shown	50	100	70.6	100	58
Numata et al[188] (predictive value for pathological LatLNM testing OSNA-MesLNM)	Not shown	None	17	100	55	61.8	28	100
Tani et al[33]	Not shown	CK19	6.9 (pOSNA)	84.6	90.9	89.1%	78.6	93.7

CEA: Carcinoembryonic antigen; CK19: Cytokeratin 19; IHC: Immunohistochemistry; LatLNM: Lateral lymph node metastasis; MesLNM: Mesorectal lymph node metastasis; NPV: Negative predictive value; OSNA: One-step nucleic acid amplification; pOSNA: Pooled one-step nucleic acid amplification; PPV: Positive predictive value.

Table 3 Differences in lymph node processing methods and upstage rates of previous reports

Ref.	Subject (patients)	OSNA method	Harvested LN, n	Harvested LN, median	Dividing method of LN	Pathological staining	Measured LN by OSNA, n	Measured LN by OSNA, median	Upstage rate (pStage I and II)
Yamamoto et al[16]	Stage 0, I (85)	cOSNA	434	N/Aa	Fourb; 4 mm over diameter of LN	HE and IHC	385	4.5	16.5% (2/16)
Güller et al[12]	Stage I, II, III (22)	cOSNA	313	30 (16–60)	Fourb; 3 mm over diameter of LN	HE and IHC	56	13 (6–24)	15.3% (2/13)
Croner et al[14]	Stage I, II (103)	cOSNA	N/Aa	N/Aa	Centerc; 6 mm over halfd; 4 mm to 6 mm diameter of LN	HE	1594	14 (1–46)	25.2% (26/103)
Vogelaar et al[15]	Stage I, II (128)	cOSNA	N/Aa	N/Aa	Fourb; 10 mm over halfd; 10 or less than 10 mm diameter of LN	HE and IHC	317	Mean 15.3 (4–40)	20.2% (20/90)
Yamamoto et al[17]	Stage I, II, III (204)	cOSNA	4324	19 (3–25)	Halfd	HE	1925	8 (2–25)	17.6% (13/74)
Aldecoa et al[133]	Stage I, II (149)	cOSNA	2483	15	Centerc	HE	1940	12	51% (76/149)
Rakislova et al[125]	Stage I, II, III (188)	cOSNA, pOSNA	cOSNA 1828, pOSNA 1992	cOSNA 17 (13-22), pOSNA 20.5 (17-27)	Centerc	HE	cOSNA 1757, pOSNA 1449	cOSNA 13 (10-18), pOSNA 18 (13-25)	cOSNA 55.4% (51/92), pOSNA 20.7% (16/77)
Brito et al[126]	Stage I, II (59)	cOSNA	1046	13 (9-19)	Centerc; 5 mm over halfd; 4 or less diameter of LN	HE	753	12 (7-16)	28.8% (17/59)
Itabashi et al[18]	Stage I, II, III (195)	cOSNA	Not shown	19 (1-75)	Halfd; 4 mm over diameter of LN	HE	Not shown	8 (2-25)	15.7% (11/70) in stage II patients
Tani et al[33]	Stage II, IIIA (92)	pOSNA	2236	24.3 (5–66)	Halfd; 4 mm over diameter of LN	HE	636	6.9 (1–35)	9.1% (6/66)

^aNot applicable.

^bFour section method: dividing lymph nodes into four equal sections and sending two of these sections (50%) for pathology and one-step nucleic acid amplification (OSNA) measurement.

^cCenter-cut method: 1 mm from the center of lymph nodes are sent for pathological examination and the rest are used for OSNA.

^dHalf-division method divides lymph nodes in half and sends each 50% portion for pathology and OSNA.

HE: Hematoxylin and eosin; IHC: Immunohistochemistry; LN: Lymph node; OSNA: One-step nucleic acid amplification; pOSNA: Pooled one-step nucleic acid amplification.

Table 4 Studies analyzing colorectal cancer metastasis in sentinel lymph nodes with one-step nucleic acid amplification

Author	Patients (samples), n	Injected dye	Intraoperative OSNA assay	Examined SLNs, n
Vogelaar et al[15]	128 (325)	Patent blue dye V or indocyanine green	No	3.0 (median)
Marhic et al[184]	17	Blue dye	Yes	Not shown
Yeung et al[130]	16 (78)	Indocyanine green	No	4.9 (mean)
Esposito et al[127]	34 (51)	Indocyanine green	Yes	1.0 (median)

OSNA: One-step nucleic acid amplification; SLN: Sentinel lymph node.