Diagnostic Utility of TSSC3 and RB1 Immunohistochemistry in Hydatidiform Mole

Round 1

Reviewer 1 Report

In this paper authors conducted a study on three gestational diseases : Complete mole (CM),– Partial mole(PM) and Non-molar abortus(NMA)  using antibodies of paternal-imprinted (RB1, TSSC3, and DOG1) and maternal-imprinted (DNMT1 and GATA3) genes and established TSSC3 and RB1 could serve as a useful adjunct to  p57 for the discrimination of complete mole from partial mole and non-molar abortus.  

Based on their data few questions need to answer.

Was the three patients showing positivity to RB1 in CM also positive to TSSC3?

Did you conduct any correlation study between the level of β-hCG and TSSC3 and RB1 in all patients and if yes was there any?

Author Response

Reviewer 1

In this paper authors conducted a study on three gestational diseases: Complete mole (CM),– Partial mole (PM) and Non-molar abortus (NMA) using antibodies of paternal-imprinted (RB1, TSSC3, and DOG1) and maternal-imprinted (DNMT1 and GATA3) genes and established TSSC3 and RB1 could serve as a useful adjunct to p57 for the discrimination of complete mole from partial mole and non-molar abortus.  

Based on their data few questions need to answer.

1) Was the three patients showing positivity to RB1 in CM also positive to TSSC3?

Response: Thank you for the comments. 

Of the 3 cases of CM with RB1 positivity, 2 of them were negative toward TSSC3 and one was positive. See supplementary table 2.

 

No.	Sample No.	βhCG (mIU/mL)	Diagnosis	TSSC3		RB1
				Intensity	%	Intensity	%
1	CM001	170,624	CM	-	0	1+	30
2	CM004	583,933	CM	1+	100	1+	30
3	CM005	170,000+	CM	-	0	1+	30
4	CM006	NA	CM	-	0	2+	20
5	CM007	67,899.50	CM	-	0	-	0
6	CM008	186,596	CM	-	0	2+	30
7	CM009	542,643	CM	-	0	2+	30
8	CM010	670,773.80	CM	-	0	2+	10
9	CM011	1,000,000	CM	1+	20	2+	40
10	CM012	200,382	CM	-	0	2+	30
11	CM013	25,000	CM	-	0	2+	30
12	CM014	34,985.90	CM	-	0	2+	33
13	CM016	187,479.50	CM	-	0	2+	20
14	CM017	NA	CM	1+	20	2+	20
15	CM018	NA	CM	-	0	1+	30
16	CM019	NA	CM	-	0	2+	100
17	CM020	NA	CM	-	0	2+	30
18	PM002	184,108.50	CM	3+	20	2+	50
19	PM003	181,113.20	CM	2+	50	2+	40
20	PM005	NA	CM	1+	10	2+	30
21	PM006	NA	CM	1+	10	1+	10
22	PM007	400,000	CM	1+	30	2+	20
23	PM011	NA	CM	-	0	2+	10
24	PM012	79,677	CM	-	0	2+	30
25	PM014	89,116	CM	2+	80	2+	10
26	PM015	131,942	CM	-	0	2+	40
27	PM016	79,284	CM	-	0	2+	30
28	PM021	NA	CM	-	0	2+	10
29	PM022	NA	CM	-	0	2+	60

2) Did you conduct any correlation study between the level of β-hCG and TSSC3 and RB1 in all patients and if yes was there any?

Response: Thank you for pointing out this.

There were a total of 19 cases of CM where the β-hCG levels were available, while PM and NMA both have 9 and 4 cases with data on β-hCG.

 

Of the 19 cases of CM, 6 were positive for TSSC3 and 13 cases were negative. The average level of β-hCG in positive and negative TSSC3 expressions were 406378 and 198107, respectively. There is no statistical difference in β-hCG between positive and negative TSSC3 (p = 0.1116, t-test). See table below. 

 

As for RB1, there was only one positive case in CM, where the β-hCG was available, and 18 were negative. The average level of β-hCG in positive and negative RB1 expressions were 184108 and 272438, respectively. Unable to perform a statistical analysis as there was only one case of CM.

Complete mole: β-hCG in TSSC3

b-hCG	TSSC3 Positive	TSSC3 Negative
Mean	406378	198107
SD	341981	202074
SEM	139613.2	56045.2
N	6	13
		P = 0.1116

 

Complete mole: β-hCG in RB1

b-hCG	RB1 Positive	RB1 Negative
Mean	184,108.50	272,438
N	1	18

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript "Diagnostic utility of TSSC3 and RB1 immunohistochemistry in hydatidiform mole" is an interesting manuscript on the role of TSSC3 and RB1 immunohistochemistry in hydatidiform mole. The work is original and well structured, giving important novelty to scientific literature. The design of the project is appropriate and the results are significant. The number of patients is not so high, but, considering the rare frequency of this clinical condition, it is appropriate. The statistical analysis is well conducted and the language is acceptable.
It represents a valid work and it gives the opportunity to focus attention on a better diagnostic strategy for an early diagnosis of hydatidiform mole.

 

Author Response

Response: Thank you for the comments.

Author Response File: Author Response.docx

Reviewer 3 Report

the manuscript is interesting, well illustrated and generally well written. The manuscript can be accetped in the present form.

Author Response

Response: Thank you for the comments.

Author Response File: Author Response.docx